Methods of treating ophthalmic diseases using NPR-B agonists

ABSTRACT

Disclosed are novel compounds having NPR-B agonistic activity. Preferred compounds are linear peptides containing 8-13 conventional or non-conventional L- or D-amino acid residues connected to one another via peptide bonds.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.14/011,879, filed Aug. 28, 2013, and which issued as U.S. Pat. No.9,187,525 on Nov. 17, 2015, which is a divisional application of U.S.application Ser. No. 12/888,574, filed Sep. 23, 2010, and which issuedas U.S. Pat. No. 8,551,938 on Oct. 8, 2013, which claims priority toU.S. provisional application Ser. No. 61/287,773, filed Dec. 18, 2009,each of which is incorporated by reference in its entirety.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted electronically in ASCII format and is hereby incorporated byreference in its entirety. Said ASCII copy, created on Dec. 21, 2016, isnamed SHR-1138US3C_SL.txt and is 368,499 bytes in size.

1. Field of the Invention

The present invention generally relates to novel compounds which arcuseful in the treatment and prevention of disorders mediated bynatriuretic peptides or proteins. More particularly, the presentinvention relates to novel peptides, pharmaceutical compositionscomprising one or more novel peptides described herein, and their use inmethods of treating or preventing ocular disorders, such as glaucoma,ocular hypertension, and optic neuropathies, cardiovascular disease,kidney disease, lung disease, and other disorders mediated bynatriuretic peptides or proteins.

2. Description of Related Art

The natriuretic peptides (NP's) are a family of cyclic peptide hormonesthat have first been described by their involvement in the regulation ofnatriuresis, diuresis and blood pressure control. To date, fournatriuretic peptides have been discovered in man, i.e. atrialnatriuretic peptide (ANP; SEQ ID NO:1), B-type or brain natriureticpeptide (BNP; SEQ ID NO;2), C-type natriuretic peptide (CNP; SEQ IDNO:3) and urodilatin (SEQ ID NO:4) (see FIG. 1; and Cho et al., 1999,Heart Dis. 1:305-328). All NP's are synthesized as prepro-hormones whichare activated by proteolytic cleavage before their release into thecirculation. The NP's bind to natriuretic peptide receptors (NPR), agroup of 3 different membrane bound receptors with guanylyl cyclaseactivity (Pandey 2005, Peptides 26:901-932).

ANP was first discovered as a blood pressure decreasing factor in ratatrial homogenates in 1981 (de Bold 1981, Life Sci 28:89-94). Humanpre-pro-ANP (SEQ ID NO: 5) contains 151 amino acids and is stored afterN-terminal cleavage as 126 amino acid pro-ANP (SEQ ID NO:6),predominantly in atrial granules. Cardiac stretch, due to systemicvolume overload induces the rapid release of ANP from these stores. Uponsecretion into the circulation, the C-terminal part of pro-ANP iscleaved by the atrial peptidase corin to the biologically active 28amino acid form of ANP (SEQ ID NO:1) (Yan 2000, Proc Natl Acad Sci97:8525-8529). The remaining N-terminal part can be further cleaved into3 different hormones. i.e. Long Acting Natriuretic Peptide (LANP, aminoacids 1-30; SEQ ID NO:7), Vessel Dilator (VSDL, amino acids 31-67; SEQID NO:8) and Kaliuretic Peptide (KP, amino acids 79-98; SEQ ID NO:9)(Vesely 2004, Eur J Clin Invest 34:674-682).

After BNP was discovered in porcine brain as a factor that showed smoothmuscle relaxing activity (Sudoh T, 1988, Nature 332:78), a much greatertissue expression was found in preparations of cardiac ventricles(Mukoyama 1991, J Clin Invest 87:1402-1412), which led to the conclusionthat BNP is, similarly to ANP, a cardiac peptide hormone. Although BNPcan be found in storage granules in the atria, the expression inventricles is transcriptionally regulated (Tamura 2000, Proc Natl AcadSci 93:4239-4244). Synthesis of pre-pro-BNP is induced through cardiacwall stretch and leads to a 134 amino acid long peptide (SEQ ID NO:10)which is further cleaved by an unknown protease to yield the 108 aminoacid long pro-BNP (SEQ ID NO:11). Additional cleavage liberates theactive 32 amino acid C-terminal fragment of BNP (SEQ ID NO:2) and theinactive 76 amino acid N-terminal fragment also referred to asNT-pro-BNP (SEQ ID NO:12). To date, no known splice variants of humanBNP exists.

CNP was first isolated from porcine brain almost 10 years after thediscovery of ANP (Sudoh 1990, Biochem Biophys Res Comm 168:863-870). Itis primarily expressed in the central nervous system and endothelialcells. Unlike other NP's, CNP is nearly not present in cardiac tissue,which suggest a more paracrine function on vascular tone and muscle cellgrowth. The 126 amino acid precursor molecule pro-CNP (SEQ ID NO: 13) isprocessed by the intracellular endoprotease furin into the mature 53amino acid peptide CNP-53 (SEQ ID NO:14), which is the most abundantform in the brain (Totsune 1994, Peptides 15:37-40), endothelial cells(Stingo, 1992, Am J Phys 263:H1318-H1321) and the heart (Minamino 1991,Biochem Biophys Res Comm 179:535-542). In both, cerebral spinal fluid(Togashi 1992, Clin Chem 38:2136-2139) and human plasma (Stingo 1992, AmJ Phys 263:H1318-H1321) the most common form is CNP-22 (SEQ ID NO:3),which is generated from CNP-53 by an unknown extracellular protease.Unlike the other NP's CNP-22 lacks the C-terminal extension of the 17amino acid ring (see FIG. 1).

ANP (SEQ ID NO:1), BNP (SEQ ID NO:2) and CNP (SEQ ID NO:3) show a highlyconserved amino acid sequence among different vertebrate species (seeFIG. 1; and Cho 1999, Heart Dis. 1:305-328). The NP's are inactivated bytwo distinct mechanisms, i.e. enzymatic cleavage through neutralendopeptidases and binding to the NP clearance receptor (NPR-C; SEQ IDNO:15), which is followed by internalization and intracellulardegradation of the NP (Stoupakis 2003, Heart Dis. 5:215-223).

The discovery of the natriuretic peptides ANP, BNP and CNP was followedby the description and cloning of their specific receptors, natriureticpeptide receptor -A, -B and -C (NPR-A, -B, -C) (Fuller 1988, J BiolChem. 263:9395-9401; Chang 1989 Nature 341:68-72; Chinkers 1989, Nature338:78-83). NPR-A (SEQ ID NO:16) preferentially binds ANP and BNP, whileNPR-B (SEQ ID NO:17) is most specific for CNP and NPR-C (SEQ ID NO:15)binds all natriuretic peptides (Koller 1991, Science 252:120-123).

The primary structure of NPR-A and NPR-B contain an extracellular ligandbinding domain, transmembrane domain, intracellular kinase homologydomain containing phosphorylation sites and a C-terminal guanylatecyclase domain (reviewed in Misono 2005, Peptides 26:957-68). The latterclassifies NPR-A and NPR-B as particulate guanylate cyclases, also knownas GC-A and GC-B (E.C.4.6.1.2). In contrast, NPR-C is lackingintracellular homology domains, but evidence is increasing for NPR-C'srole not only as a scavenger receptor for natriuretic peptides, but forits' functional coupling to inhibitory G-proteins and phosphoinositideturnover (Maack 1987, Science 238:675-678; Murthy and Makhlouf 1999, JBiol Chem 274:17587-17592; Anand-Srivastava 2005, Peptides26:1044-1059). Reflecting the grade of sequence homology in natriureticpeptides, natriuretic peptide receptors show a high degree of homologyin their extracellular ligand binding domains, with the calculatedsimilarities being 41% between NPR-A and NPR-B and 29% between NPR-A andNPR-C (van den Akker 2001, J Mol Biol. 311:923-937).

Ligand binding to NPRs requires a dimer of glycosylated receptorsubunits (Fenrick et al. 1994, Mol Cell Biochem. 137:173-182; Kuhn 2003,Circ Res. 93:700-709) and is followed by a conformational change leadingto activation of the guanylate cyclase domains. Subsequently, activityof particulate guanylate cyclases is regulated through phosphorylation(reviewed in Kuhn 2003, Circ Res. 93:700-709). Phosphorylation of NPRsis maximal in the basal state, while ligand binding is followed bydephosphorylation and subsequent desensitization of the receptor.

Natriuretic receptors are expressed in many tissues throughout theorganism. NPR-A, NPR-B and NPR-C are present in the cardiovascularsystem and the kidney, with NPR-C being the most abundant receptorsubtype accounting for 80% of NPR-expression in some tissues. NPR-B ispresent in a particularly high level in rat pineal gland, testis andovaries. NPR-A and NPR-B ligands both induce endothelium-independentvasorelaxation, where ANP and BNP mainly act on arterial vasculature. Incontrast, CNP mainly targets the venous system, with the exception ofcoronary arteries, that relax in response to CNP stimulation (Marton etal. 2005, Vascul Pharmacol 43:207-212). Importantly, induction ofhypotension via NPR-B activation requires 10-fold higher concentrationsof ligand compared to blood pressure reduction in response to NPR-Aactivation (Wei et al. 1993, Am J Physiol. 264:H71-H73; Woods and Jones1999, Am J Physiol. 276:R1443-R1452). Relaxation of smooth muscle byactivation of NPR-B has been shown in a variety of tissues, includingblood vessels, seminiferous tubules and uterus. Also contraction of theocular trabecular meshwork tissue is reduced by activation ofnatriuretic peptide receptors, confirming functional similarities oftrabecular meshwork and smooth muscle cells (Stumpff and Wiederholt2000, Ophthalmologica 214:33-53).

Another main target organ of natriuretic peptides is the kidney. Ligandsof NPR-A induce natriuresis and diuresis by a dual mechanism (reviewedin Beltowski and Wojcicka 2002, Med Sci Monit. 8:RA39-RA52): (1)increased excretion of sodium by a reduced re-uptake of sodium ions inthe distal tubulus, subsequently leading also to higher retention ofwater in the final urine; and (2) dilation of the affluent andconcomitant contraction of the effluent glomerular capillary, increasingglomerular filtration rate, at the cost of reduction of renal perfusion(Endlich and Steinhausen 1997, Kidney Int. 52:202-207). In contrast toNPR-A-specific ligands, NPR-B-specific ligands do not induce significantnatri- and diuresis, and in addition, show a peculiarity regardingglomerular flow regulation: CNP was shown to dilate both affluent andeffluent capillaries in the glomerulus, thus increasing renal bloodflow, but not glomerular filtration (Endlich and Steinhausen 1997,Kidney Int. 52:202-207).

In addition to effects of NP-receptor (NPR) activation on blood pressureand kidney function, powerful effects of natriuretic peptides onproliferative processes in a variety of cell types have been documentedin the literature. Antiproliferative properties of NPR activation aredocumented for vascular smooth muscle cells, fibroblasts of differentorigins, mesangial cells, cancer cells and chondrocytes (reviewed inSchulz 2005, Peptides 26:1024-1034). At least for VSMC, evidence for theinvolvement of the transcription factor GAX in the regulation ofproliferation has given an indication as to which intracellularmechanisms might be involved in growth regulation through NPR (Yamashitaet al. 1997, Hypertension 29:381-387). Though tissue growth is mainlyregulated by proliferative activity, some organs feature variations incell size to influence tissue mass. This might be a physiologicalprocess, as during endochondral ossification, when chondrocytes matureby undergoing hypertrophy, or a pathological event, as in cardiachypertrophy, which often precedes chronic heart failure. Both of theabove-mentioned events of hypertrophy are regulated by NPR-B. NPR-Bdeficiency causes dwarfism due to abnormal endochondral ossification,characterized by size reduction of the hypertrophic zone of theepiphyseal growth plate (Bartels et al. 2004, Am J Hum Genet. 75:27-34;Tamura et al. 2004, Proc Natl Acad Sci. 101:17300-17305).

Quite different, a partial knock out of NPR-B in rats promoted cardiachypertrophy, i.e. hypertrophy of cardiomyocytes (Langenickel et al.2006, Proc Natl Acad Sci. 103:4735-4740).

Natriuretic peptides, having activity at the natriuretic receptors, werelater discovered in various tissues, as well. For example, ANP wasdiscovered in the early 1980s as an endogenous diuretic and vasorelaxantpeptide, whose principle circulating form consists of 28 amino acids(SEQ ID NO:1). Subsequently, other natriuretic peptides, such as BNP(SEQ ID NO:2) and CNP (SEQ ID NO:3), were discovered. The presence ofnatriuretic peptides and their receptors in ocular tissues, especiallythose involved in the regulation of IOP, have been demonstrated. Forexample, in rat and rabbit eyes, ANP, BNP, and CNP, as well as NPR-A,NPR-B, and NPR-C mRNA were found in the ciliary processes, retina, andchoroid (Mittag et al. 1987, Curr Eye Res. 6:1189-1196; Nathanson 1987,Invest Ophthalmol Vis Sci. 28:1357-1364; Fernandez-Durango et al. 1995,Exp Eye Res. 61:723-729). Similar results were found in bovine ciliaryprocesses and cultured bovine ciliary epithelial cells. (Millar et al.1997, J Ocul Pharmacol Ther. 13:1-11; Shahidullah and Wilson 1999, Br JPharmacol. 127:1438-1446). The presence of the peptides and theirreceptors in the ciliary epithelium suggests that they may play a rolein the production of aqueous humor.

In addition to the ciliary processes, natriuretic peptide receptors werealso found in tissues associated with the outflow of aqueous humor. ANPbinding sites were localized in the longitudinal ciliary muscle of theguinea pig. (Mantyh et al. 1986, Hypertension. 8:712-721). In culturedhuman TM and ciliary muscle cells, CNP is the most potent andefficacious in stimulating the production of cyclic GMP, indicating thepresence of functional NPR-B. Activation of this receptor reducescarbachol-induced calcium influx. (Pang et al. 1996, Invest OphthalmolVis Sci. 37:1724-1731). This result predicts that activation of NPR-Bshould cause relaxation of these tissues. Indeed, CNP significantlydecreases the carbachol-induced contraction of monkey and human ciliarymuscles. (Ding and Abdel-Latif, 1997, Invest Ophthalmol Vis Sci.38:2629-2638). Change in contractility in TM and ciliary muscle mayaffect the outflow facility of aqueous humor.

Cyclic GMP and compounds that increase cyclic GMP in ocular tissues,such as nitric oxide donors, have been shown to lower IOP. (Nathanson1988, Eur J Pharmacol. 147:155-156; Becker 1990, Invest Ophthalmol VisSci. 31:1647-1649; Nathanson 1992, J Pharmacol Exp Ther. 260:956-965;Stein and Clack 1994, Invest Ophthalmol Vis Sci. 35:2765-2768). Sincenatriuretic peptides potently increase cyclic GMP production, they werepredicted to lower IOP, too. In the past 20 years, the natriureticpeptides have been shown to be highly effective as IOP-lowering agents.For example, various researchers have independently shown thatintravitreal injection of ANP in rabbits consistently and significantlylowers IOP. This effect lasts for many hours. (Sugrue and Viader, 1986,Eur J Pharmacol. 130:349-350; Mittag et al. 1987, Curr Eye Res.6:1189-1196; Nathanson 1987 Invest Ophthalmol Vis Sci. 28:1357-1364;Korenfeld and Becker 1989, Invest Ophthalmol Vis Sci. 30:2385-2392;Takashima et al. 1996, Invest Ophthalmol Vis Sci. 37:2671-2677). The IOPeffect of ANP correlates with an increase in cyclic GMP production inthe iris-ciliary body. (Korenfeld and Becker 1989, Invest Ophthalmol VisSci. 30:2385-2392). Intravitreal injection of BNP (Takashima et al.1996, Invest Ophthalmol Vis Sci. 37:2671-2677) or CNP (Takashima et al.1998, Exp Eye Res. 66:89-96) is also highly efficacious in lowering IOP.In addition to intravitreal injection, subconjunctival (Yang et al.1997, Chin J Ophthalmol. 33:149-151) or intracameral (Sugrue and Viader1986, Eur J Pharmacol. 130:349-350; Fernandez-Durango et al. 1999, Eur JPharmacol. 364:107-113) injection of the natriuretic peptides have beenshown to be ocular hypotensive as well. Systemic administration of ANPin the rabbit, (Tsukahara et al. 1988, Ophthalmologica. 197:104-109) orhuman (Diestelhorst and Krieglstein 1989, Int Ophthalmol. 13:99-101)also lowers IOP. Unfortunately, it has not been possible to deliverthese peptides topically due to their inability to penetrate the cornea.Therefore, these potent and efficacious IOP-lowering compounds have notbeen developed for such use.

There is a need for novel NPR-B agonists having improvedbioavailability, as compared to isolated or synthesized natriureticpeptides, that can be used in the treatment of natriureticpeptide-mediated disorders, such as ocular disorders, diabetes-relateddisorders, vascular disorders, cardiac and cardiovascular pathologies,inflammation and other disorders described herein. The novel NPR-Bagonists, compositions and methods of the present invention meet theseneeds.

SUMMARY OF THE INVENTION

The present invention provides novel NPR-B agonists, also referred toherein as natriuretic peptide mimics or similars, that aretherapeutically useful for lowering intraocular pressure (IOP) andtreating other disorders where activation of the type B natriureticpeptide receptor will be beneficial. Specifically, the inventionprovides novel NPR-B agonists that activate the type B natriureticpeptide receptor (NPR-B). The invention further provides compositionscontaining such novel NPR-B agonists. The compositions provided hereinmay be ophthalmic compositions for use in methods of treating orpreventing particular ophthalmic diseases such as glaucoma, preferablyby lowering intraocular pressure, using such novel NPR-B agonists.Alternatively, the compositions provided herein may be used in methodsof treating or preventing cardiovascular disorders, kidney disease, lungdisease, skeletal disorders, infertility, and other disorders mediatedby natriuretic peptides or proteins.

The invention is in part based on the inventors' finding that the novelNPR-B agonists described herein can provide improved bioavailability,increased chemical stability, and increased metabolic stability in bodyfluids or tissues, due to their significantly reduced molecular size ascompared to the known natriuretic peptides. Certain embodiments of thepresent application generally pertain to novel peptides containingmodified amino acids and that bind to and activate NPR-B with highspecificity, as described in more detail herein.

It is specifically contemplated that any limitation discussed withrespect to one embodiment of the invention may apply to any otherembodiment of the invention. Furthermore, any composition of theinvention may be used in any method of the invention, and any method ofthe invention may be used to produce or to utilize any composition ofthe invention.

As used herein, the term “NPR-B agonist” refers to the novel moleculesdescribed herein that activate the NPR-B with high potency.

The use of the term “or” in the claims is used to mean “and/or” unlessexplicitly indicated to refer to alternatives only or the alternativeare mutually exclusive, although the disclosure supports a definitionthat refers to only alternatives and “and/or.”

Throughout this application, the term “about” is used to indicate that avalue includes the standard deviation of error for the device and/ormethod being employed to determine the value.

As used herein the specification, “a” or “an” may mean one or more,unless clearly indicated otherwise. As used herein in the claim(s), whenused in conjunction with the word “comprising,” the words “a” or “an”may mean one or more than one. As used herein “another” may mean atleast a second or more.

Other objects, features and advantages of the present invention willbecome apparent from the following detailed description. It should beunderstood, however, that the detailed description and the specificexamples, while indicating preferred embodiments of the invention, aregiven by way of illustration only, since various changes andmodifications within the spirit and scope of the invention will becomeapparent to those skilled in the art from this detailed description.

BRIEF DESCRIPTION OF THE FIGURES

The following figures form part of the present specification and areincluded to further demonstrate certain aspects of the presentinvention. The invention may be better understood by reference to one ormore of these drawings in combination with the detailed description ofspecific embodiments presented herein.

FIG. 1. Illustrates the amino acid sequence of ANP (SEQ ID NO;1), BNP(SEQ ID NO:2) and CNP (SEQ ID NO:3).

FIG. 2. Illustrates the effects of CNP, ANP, BNP and mini-ANP (SEQ IDNO:18) on cyclic GMP production in GTM-3 cells. GTM-3 cells have beenshown to express NPR-B (Pang et al. 1996, Invest Ophthalmol Vis Sci.37:1724-1731). The cells were treated with CNP (triangles), ANP(squares), BNP (diamonds) and mini-ANP (circles). The symbols representmean values and standard deviations. The highest concentration ofcompounds used was 45 μM for ANP, BNP and mini-ANP and 5 μM for CNP.EC50 values were determined using the 4-Parameter Logistic Equation. CNPEC50=38.8 nM, ANP EC50=1.63 μM, BNP EC50=1.18 μM, mini-ANP EC50>45 μM.The Emax (maximum activation) of each compound was determined relativeto the maximum activation of CNP, i.e. CNP Emax=100%, ANP Emax=15%, BNPEmax=20% and mini-ANP Emax=0%.

FIG. 3. Illustrates the effects of CNP, ANP, BNP and mini-ANP on cyclicGMP production in NPR-A transfcctcd 293-T cells. NPR-A transfcctcd 293-Tcells were treated with CNP (triangles), ANP (squares), BNP (diamonds),and mini-ANP (circles). The symbols represent mean values and standarddeviations. EC₅₀ was determined using the 4-Parameter Logistic Equation.EC₅₀ of ANP=73.0 nM, EC₅₀ of CNP=1.60 μM, EC₅₀ of BNP=1.85 μM, EC₅₀ ofmini-ANP=1.54 μM.

DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

The present invention is in part based on the finding that novel NPR-Bagonists having improved bioavailability as compared to knownnatriuretic peptides are useful for lowering elevated intraocularpressure and treating glaucoma. Thus, the present invention is generallydirected to novel NPR-B agonists and their use in methods of treating orpreventing disorders mediated by natriuretic peptides or proteins. Inone particularly preferred embodiment, the novel NPR-B agonistsdescribed herein are formulated for the treatment of ophthalmic diseasessuch as glaucoma, preferably by lowering the elevated intraocularpressure often associated with glaucoma, using a pharmaceuticalcomposition that comprises one or more novel NPR-B agonists, asdescribed herein. In other preferred embodiments, the novel NPR-Bagonists described herein are formulated for the treatment of othernatriuretic peptide- or protein-mediated disorders such ascardiovascular disorders, kidney disorders, lung disorders, skeletaldisorders, fertility disorders, and fibrosis.

The hallmark feature of all known NP's is the 17 amino acid ring whichis formed by an intramolecular cysteine bridge (see FIG. 1). Theintegrity of the cyclic structure of NP's is believed to be critical forthe functional activity, i.e. NP receptor transduced cGMP production.The present inventors have discovered that certain linear peptides, suchas the novel peptides described herein, having increased chemical andmetabolic stability and the improved bioavailability as compared toknown NP's, are useful in the treatment of natriuretic peptide- orprotein-mediated disorders.

A. Novel Peptides

The present invention provides novel NPR-B agonists having biologicalactivity that is improved in certain aspects as compared to that of theknown natriuretic peptides. The novel peptides of the invention includeconventional and non-conventional amino acids. Conventional amino acidsare identified according to their standard, three-letter codes, as setforth in Table 1, below.

TABLE 1 For conventional amino acids the 3-letter codes were used:3-letter codes Amino acids 3-letter codes Amino acids Ala Alanine MetMethionine Cys Cysteine Asn Asparagine Asp Aspartic acid Pro Proline GluGlutamic acid Gln Glutamine Phe Phenylalanine Arg Arginine Gly GlycineSer Serine His Histidine Thr Threonine Ile Isoleucine Val Valine LysLysine Trp Tryptophane Leu Leucine Tyr Tyrosine

Non-conventional amino acids are identified according to a three-lettercode, or other abbreviation, when present in the novel NPR-B agonists ofthe invention. Table 2, below, provides the full name, three-letter codeor abbreviation, and structure of each non-conventional amino acidappearing in the sequences of the novel peptides described herein.

TABLE 2 List of abbreviations of non-conventional amino acids and otherchemical structures. Name Abbr Structure(S)-2-((S)-3-amino-2,5-dioxopyrrolidin-1-yl)-5-guanidinopentanoic acidDim-Arg

rac-2-amino-4-morpholinobutanoic acid AR-385-017

(S)-2-amino-3-(2H-tetrazol-5-yl) propanoic acid AR-314-145

rac (1S,2S)-2-(octylcarbamoyl) cyclohexane carboxylic acid AR-314-171

rac (1S,2S)-2-(hexylcarbamoyl) cyclohexane carboxylic acid AR-314-170

rac (1R,2S)-2-octylcarbamoyl) cyclohexane carboxylic acid AR-314-169

(S)-2-(6-hexanamido-1- oxoisoindolin-2-yl)-3- phenylpropanoic acidAR-385-008

(S)-2-(4-octanamido-1,3- dioxoisoindolin-2-yl)-3- phenylpropanoic acidAR-314-172

(S)-2-(5-hexanamido-1,3- dioxoisoindolin-2-yl)-3- phenylpropanoic acidAR-385-042

(S,S)-2-(3-methyl-3-octanoylamino- 2-oxo-pyrrolidin-1-yl)-3-phenyl-propionic acid AR-314-102

2-(7-Octanoyl-1-oxo-2,7-diaza- spiro[4.5]dec-2-yl)-3-phenyl- propionicacid acid AR-314-087

1-(3-Methyl-butyl)-piperazine AR-201-124

Cycloheptyl-pyrrolidin-2-ylmethyl- amine ES-283-049

(S)-Amino-thiophen-2-yl-acetic acid BB727

(R)-Amino-thiophen-2-yl-acetic acid BB726

2-Octylsulfanyl-propionic acid AR-201-073

5-Pentylsulfanylmethyl-oxazole-2- carboxylic acid AR-201-072

4-(4-Butyl-thiazol-2-ylamino)- benzoic acid AR-201-069

4-(5-Butyl-thiazol-2-ylamino)- benzoic acid AR-201-068

2-Hexylamino-oxazole-4-carboxylic acid AR-201-062

2-Hexanoylamino-oxazole-4- carboxylic acid AR-201-059

3-Hexyloxy-isoxazole-5-carboxylic acid AR-201-058

2-Hexanoylamino-isonicotinic acid AR-201-054

Octanoic acid 1-carboxy-ethyl ester AR-201-049

Dodecanoic acid 1-carboxy-2- phenyl-ethyl ester AR-201-048

(R)-2-Amino-4-(piperidin-1-yl) butanoic acid abu(pip)

8-amino-3,6-dioxaoctanoic acid Adx

(2,3,4,5,6-Pentahydroxy- hexylidenaminooxy)-acetic acid Gluc-Aoa

5-((4S)-2-oxohexahydro-1H- thieno[3,4-d]imidazol-4- yl)pentanoic acid 74

Adamantan-2-yl-amine 504

Cyclohexylamine 558

Cyclopentylamine 559

2-((1S,2R,4R)-bicyclo[2.2.1]heptan- 2-yl)acetic acid 779

2-Phenethyl-benzoic acid 785

Dodecanoic acid 832

Aniline 873

Octanesulfonyl chloride 933

Hexyl chloroformate 1270

3-Phenyl-propionic acid 1281

4-Phenyl-butyric acid 1319

5-Phenyl-pentanoic acid 1320

4-Cyclohexyl-butyric acid 1339

3-Cyclohexyl-propionic acid 1340

(S)-3,3-dimethylbutan-2-amine 1381

2-(hexylamino)acetic acid 1625-Ac

Piperidine-1,2-dicarboxylic acid 1-benzyl ester 1695

4-Methyl-cyclohexyl-amine 1859

(1R,2R)-2-methylcyclohexanamine 1860

2-(2-Methoxy-ethoxy)-ethoxy]- acetic acid 1888

(1R,2R,4R)-bicyclo[2.2.1]heptan-2- amine 1906

(2-Methoxy-ethoxy)-acetyl chloride 1913

(1R,2R)-2-(benzyloxy) cyclohexanamine 1934

(S)-1,2,3,4-tetrahydronaphthalen-1- amine 2118

(S)-3-methylpiperidine 2137

4-(4-Methoxy-phenyl)-butyric acid 2553

(1R,2R,4R)-1,7,7- trimethylbicyclo[2.2.1]heptan-2- amine 2797

2-((2S,3R,4R,5R)-2,3,4,5,6- pentahydroxyhexylamino)acetic acid 2857-Ac

Cyclobutyl-amine 2906

(S)-2-cyclopentylhexanoic acid 3218

3-Amino-4-hydroxy-benzoic acid 3421

1-Ethyl-propyl-amine 3791

(R)-2-methylbutan-1-amine 3806

2-Ethyl-butyl-amine 3816

3-(4-Bromo-phenyl)-propionic acid 4703

(4-Butoxy-phenyl)-acetic acid 4734

(1S,2R)-2- aminocyclohexanecarboxamide 5116

(1R,2S)-ethyl 2- aminocyclohexanecarboxylate 5118

(1R,2R)-ethyl 2- aminocyclohexanecarboxylate 5119

1-Propyl-butyl-amine 5121

(S)-3-amino-1-ethylazepan-2-one 5164

Decanoic acid 5587

(2-Butoxy-ethoxy)-acetic acid 6013

(E)-dodec-2-enoic acid 6014

(Z)-dodec-5-enoic acid 6015

(2S)-2-octylcyclopropanecarboxylic acid 6056

3-Octylsulfanyl-propionic acid 6057

7-Butylsulfanyl-heptanoic acid 6058

3-(Octane-1-sulfinyl)-propionic acid 6059

3-(Octane-1-sulfonyl)-propionic acid 6059(O)

rac-6-Hydroxy-decanoic acid (6071-OH)

rac-7-Hydroxy-dodecanoic acid (6072-OH)

5-Butyl-2H-pyrrazole-3-carboxylic acid 6182

2-Pentyl-benzooxazole-5-carboxylic acid 6988

(R)-2-aminobutanoic acid abu

3-Amino-1-carboxymethyl-pyridin- 2-one Acp

(S)-2-((S)-3-amino-2-oxopyrrolidin- 1-yl)-3-phenylpropanoic acid AFL

(S)-2-((R)-3-amino-2-oxopyrrolidin- 1-yl)-3-phenylpropanoic acid aFL

(R)-2-((R)-3-amino-2-oxopyrrolidin-1-yl)-3-phenylpropanoic acid afL

2-Aminoisobutyric acid Aib

2-Aminoindan-2-carboxylic acid Aic

rac-α-Methyl-leucine Ami

(R)-α-methyl-proline Amp

1-Aminomethyl- cyclopropanccarboxylic acid Amcp

4-Amino-piperdine-4-carboxylic acid Apc

4-Amino-1-(2-amino-ethyl)- piperidine-4-carboxylic acid Apc(Ac)

4-Amino-1-ethyl-piperidine-4- carboxylic acid Apc(Et)

4-Amino-1-methyl-piperidine-4- carboxylic acid Apc(Me)

(2S,4S)-4-aminopyrrolidine-2- carboxylic acid Apr

Azetidine-3-carboxylic acid Az3

(S)-azetidin-2-carboxylic acid Aze

(R)-azetidin-2-carboxylic acid aze

β-Alanine Bal

(S)-β-Homolysine Bhk

(2S,4R)-4-(benzyloxy)pyrrolidine-2- carboxylic acid Bhp

(R)-β-homoleucine Ble

rac-2-amino-3-phenyl-butyric acid Bmf

(S)-2-((S)-3-(carboxymethyl)-2- oxopiperazin-1-yl)-5- guanidinopentanoicacid cDR

(S)-β-cyc1ohexylalanine Cha

Cycloheptyl-amine Che

(S)-Cyclohexylglycine Chg

(2S,4S)-4-hydroxypyrrolidine-2- carboxylic acid Chy

(S)-2-amino-2-cyclopropylacctic acid Cpa

(S)-2-amino-2-cyclopentylacetic acid Cpg

rac-(3R,4S)-cis-methanoproline Cpp

(S)-2-amino-3-(tert-butylthio) propanoic acid ctb

(S)-2-Amino-3-sulfopropanoic acid Cya

(R)-2,4-diaminobutanoic acid dab

(R)-2-amino-3-(neopentylamino) propanoic acid dap(1464)

(R)-2-amino-3-(bis(2-aminoethyl) amino)propanoic acid dap(6263)2

(R)-2-amino-3-(bis((1H-imidazol-2- yl)methyl)amino)propanoic aciddap(3846)2

(R)-2-amino-3-(piperidin-4- ylmethylamino)propanoic acid dap(6238)

((R)-2-amino-4-(dimethylamino) butanoic acid dab(Me2)

(R)-2,3-diaminopropanoic acid dap

(S)-2-amino-3-(dimethylamino) propanoic acid Dap(Me2)

(R)-2-amino-3-(dimethylamino) propanoic acid dap(Me2)

2-Amino-2-ethyl-butyric acid Deg

2-Aminoacrylic acid Dha

(S)-2,5-dihydro-1H-pyrrole-2- carboxylic acid Dhp

(R)-2,2-dimethylthiazolidine-4- carboxylic acid Dtp

(S)-3,4-dichloro-phenylalanine Eaa

(S)-2-(3-amino-2-oxoazepan-1-yl acetic acid Eah

rac-Imidazolidine-2-carboxylic acid Eal

(S)-4-methyl-2-((S)-6-oxo-1,7- diazaspiro[4.4]nonan-7-yl)pentanoic acidEam

rac-1-amino-2,3-dihydro-1H- indene-1-carboxylic acid Eao

2,3-Dihydro-1H-indole-2-carboxylic acid Eat

(2S,4S)-4-phenylpyrrolidine-2- carboxylic acid Eay

(R)-thiazolidine-4-carboxylic acid Eaz

1-Aminocyclopropanecarboxylic acid Ebc

(R)-2-amino-3-(methylsulfanyl) propanoic acid Ebe

1-Amino-cyclopentanecarboxylic acid Eca

2-Amino-3-piperidin-4-yl-propionic acid Egg

1-aminocyclohexanecarboxylic acid Egz

(1S,3R)-3-aminocyclohexane carboxylic acid Fio

trans-4-(aminomethyl)cyclohexane carboxylic acid Fir

Amino-piperidin-3-yl-acetic acid Fhy

(S)-2-amino-2-(piperidin-4-yl)acetic acid Fhz

(2S,4S)-4-fluoropyrrolidine-2- carboxylic acid Fpr

4-aminobutyric acid Gab

(R)-2-amino-3-guanidinopropanoic acid gdp

(2S,4R)-4-guanidinopyrrolidine-2- carboxylic acid Gup

(2S,3S)-3-hydroxypyrrolidine-2- carboxylic acid H3p

Hexanoic acid Hex

(S)-homo-phenylalanine Hfe

(S)-2-aminooctanoic acid Hgl

(R)-2-aminooctanoic acid hgl

(S)-2-amino-5-methylhexanoic acid Hle

(S)-homo-serine Hse

(R)-homo-serine hse

(2S,4R)-4-hydroxypyrrolidine-2- carboxylic acid Hyp

Piperidine-4-carboxylic acid Inp

Dodecane Lau

(R)-2-amino-6-(dimethylamino) hexanoic acid lys(Me2)

3-Aminomethyl-benzoic acid Mam

(R)-2-amino-4-(methylsulfonyl) butanoic acid metO₂

(S)-meta-chloro-phenylalanine Mcf

(S)-4-hydroxy-3-Iodo-phenylalanine Miy

(S)-meta-methyl-phenylalanine Mmf

(S)-3-(3-Pyridyl)-alanine Mpa

(3-Amino-phenyl)-acetic acid Mpe

(S)-meta-trifluoromethyl- phenylalanine Mtf

(R)-2-amino-4-guanidinobutanoic acid nar

rac-(2,3-Dihydroxy-propylamino)- acetic acid Nbhp

4-Butyl-thiazole Nbt

(3-Hydroxy-propylamino)acetic acid Nhpr

Phenethylamino-acetic acid NHfe

(S)-para-nitro-phenylalanine Nif

rac-Nipecotic acid Nip

(S)-Norleucine Nle

(R)-Norleucine nle

(S)-N-methyl-alanine Nma

(S)-N-methyl-aspartic acid Nmd

(S)-N-methyl-phenylalanine Nmf

(S)-N-methyl-isoleucine Nmi

(S)-N-methyl-lysine Nmk

(S)-N-methyl-leucine Nml

(S)-N-methyl-arginine Nmr

(S)-2-amino-4,4-dimethylpentanoic acid Npg

4,4-Dimethyl-2-methylamino- pentanoic acid SH-112-158

Benzylamino-acetic acid NPhe

(S)-4-methyl-2-(propylamino) pentanoic acid Npl

(S)-norvaline Nva

(R)-norvaline nva

Octanoic acid Occ

octane Oct

(2S,3aS,7aS)-octahydro-1H-indole- 2-carboxylic acid Oic

(S)-3-(2-pyridyl)-alanine Opa

(S)-ornithine Om

(R)-ornithine orn

(R)-2-amino-5-(dimethylamino) pentanoic acid orn(Me2)

(S)-ortho-trifluoro-phenylalanine Otf

Piperazin-1-yl-acctic acid Paa

(S)-para-amino-phenylalanine Paf

(4-Aminomethyl)-benzoic acid Pam

(S)-para-bromo-phenylalanine Pbf

(2S,3R)-3-aminopyrrolidine-2- carboxylic acid Pca

(S)-para-chloro-phenylalanine Pcf

(S)-para-fluoro-phenylalanine Pff

(S)-phenylglycine Phg

(S)-pipecolinic acid Pip

(R)-pipecolinic acid pip

(S)-para-methyl-phenylalanine Pmf

(S)-para-methoxy-phenylalanine Pmy

(S)-3-(4-Pyridyl)-alanine Ppa

(4-Amino-phenyl)-acetic acid Ppe

(S)-2-amino-3-(phosphonooxy) propanoic acid Psc

(2S,3R)-2-Amino-3- (phosphonooxy) butanoic acid Pth

Sarcosine Sar

5-Butyl-thiazole Sbt

(S)-nipecotic acid Sni

(2S,4R)-4-aminopyrrolidine-2- carboxylic acid Tap

(2S,4R)-4-(dimethylamino) pyrrolidine-2-carboxylic acid Tap(2Me)

(2S,4R)-4-acetamidopyrrolidine-2- carboxylic acid Tap(Ac)

(2S,4R)-4-(2-aminoethylamino) pyrrolidine-2-carboxylic acid Tap(Ae)

(2S,4R)-4-(S)-3-amino-3- carboxypropaneamido)pyrrolidine- 2-carboxylicaicd Tap(Asp(-))

4-(3-Amino-propylamino)- pyrrolidine-2-carboxylic acid Tap(Ap)

(2S,4R)-4-(3-aminopropanamido) pyrrolidine-2-carboxylic acid Tap(Bal)

(2S,4R)-4-(diethylamino)pyrrolidine-2- carboxylic acid Tap(Et2)

(2S,4R)-4-(ethylamino)pyrrolidine- 2-carboxylic acid Tap(Et)

(2S,4R)-4-(2-aminoacetamido) pyrrolidine-2-carboxylic acid Tap(G)

(S)-α-tert-butylglycine Tbg

(R)-α-tert-butylglycine tbg

(2S,4R)-4-fluoropyrrolidine-2- carboxylic acid Tfp

(S)-2-thienyl-alanine Thi

(S)-3-thienyl-alanine Thk

(S)-thiazolidinc-4-carboxylic acid Thz

(S)-1,2,3,4-tetrahydroisoquinoline- 3-carboxylic acid Tic

4-Amino-thiazol-2-carboxylic acid Tnc

(S)-2,3-Diamino-propionic acid (side chain prolongation) Udp

The novel NPR-B agonists of the invention comprise the general aminoacid sequence of Formula I:B-Xaa₁-Xaa₂-Xaa₃-Xaa₄-Xaa₅-Xaa₆-Xaa₇-Xaa₈-Xaa₉-Xaa₁₀-Z   (I)

wherein

B is selected from the group consisting of H, R^(b1)—, R^(b2)—C(O)—,R^(b2)—S(O₂)—, R^(b3)-Baa-;

Baa is a conventional α-amino acid, a non-conventional α-amino acid or aβ-amino acid;

R^(b1) is selected from C₁-C₁₂ alkyl optionally substituted byNR^(b4)R^(b5), OH, OR^(b6), C₃-C₈ cyclic alkyl, aryl, heteroaryl, orheterocyclyl; C₁-C₁₂ alkenyl optionally substituted by NR^(b4)R^(b5),OH, OR^(b6), C₃-C₈ cyclic alkyl, aryl, heteroaryl, or heterocyclyl;C₁-C₁₂ alkyl aryl optionally substituted by NR^(b4)R^(b5), OH, orOR^(b6); C₁-C₁₂ alkynyl optionally substituted by NR^(b4)R^(b5), OH,OR^(b6), C₃-C₈ cyclic alkyl, aryl, heteroaryl, or heterocyclyl; arylC₁-C₁₂ alkyl optionally substituted by NR^(b4)R^(b5), OH, OR^(b6), C₃-C₈cyclic alkyl, aryl, heteroaryl, or heterocyclyl; C₁-C₁₂ alkyl C₃-C₈cyclic alkyl optionally substituted by NR^(b4)R^(b5), OH, OR^(b6), aryl,heteroaryl, or heterocyclyl; C₃-C₆ cyclic alkyl C₁-C₁₂ alkyl optionallysubstituted by NR^(b4)R^(b5), OH, OR^(b6), aryl, heteroaryl, orheterocyclyl; C₁-C₉ alkylthio C₂-C₁₀ alkyl optionally substituted byNR^(b4)R^(b5), OH, OR^(b6), C₃-C₈ cyclic alkyl, aryl, heteroaryl, orheterocyclyl; C₁-C₉ alkylsulfonyl C₁-C₄ alkyl optionally substituted byNR^(b4)R^(b5), OH, OR^(b6), C₃-C₈ cyclic alkyl, aryl, heteroaryl, orheterocyclyl; C₁-C₉ alkylsulfoxyl C₁-C₁₀ alkyl optionally substituted byNR^(b4)R^(b5), OH, OR^(b6), C₃-C₈ cyclic alkyl, aryl, heteroaryl, orheterocyclyl; CH₃—(CH₂)_(qb)—O—[—CH₂—(CH₂)_(nb)O]_(mb)—CH₂—(CH₂)_(pb)—,2-thiazolo optionally substituted by C₁₋₈ alkyl;

qb=0-3

nb=1-3

mb=1-3

pb=1-3

R^(b2) is selected from C₁-C₁₂ alkyl optionally substituted byNR^(b4)R^(b5), OH, OR^(b6), C₃-C₈ cyclic alkyl, aryl, heteroaryl, orheterocyclyl; C₁-C₁₂ alkenyl optionally substituted by NR^(b4)R^(b5),OH, OR^(b6) C₃-C₈ cyclic alkyl, aryl, heteroaryl, or heterocyclyl; arylC₁-C₁₂ alkyl optionally substituted by NR^(b4)R^(b5), OH, OR^(b6), C₃-C₈cyclic alkyl, aryl, heteroaryl, or heterocyclyl; C₁-C₁₂ alkynyloptionally substituted by NR^(b4)R^(b5), OH, OR^(b6)C₃-C₈ cyclic alkyl,aryl, heteroaryl, or heterocyclyl; C₁-C₁₂ alkyl aryl optionallysubstituted by NR^(b4)R^(b5), OH, or OR^(b6); C₁-C₁₂ alkyl C₃-C₈ cyclicalkyl optionally substituted by NR^(b4)R^(b5), OH, OR^(b6), C₃-C₈ cyclicalkyl, aryl, heteroaryl, or heterocyclyl; C₃-C₆ cyclic alkyl C₁-C₁₂alkyl optionally substituted by NR^(b4)R^(b5), OH, OR^(b6)C₃-C₈ cyclicalkyl, aryl, heteroaryl, or heterocyclyl; C₁-C₉ alkylthio C₁-C₁₀ alkyloptionally substituted by NR^(b4)R^(b5), OH, OR^(b6), C₃-C₈ cyclicalkyl, aryl, heteroaryl, or heterocyclyl; C₁-C₉ alkylsulfonyl C₁-C₁₀alkyl optionally substituted by NR^(b4)R^(b5), OH, OR^(b6), C₃-C₈ cyclicalkyl, aryl, heteroaryl, or heterocyclyl; C₁-C₉ alkylsulfoxyl C₁-C₄alkyl optionally substituted by NR^(b4)R^(b5), OH, OR^(b6), C₃-C₈ cyclicalkyl, aryl, heteroaryl, or heterocyclyl,CH₃—(CH₂)_(qb)—O—[—CH₂—(CH₂)_(nb)O]_(mb)—CH₂—(CH₂)_(pb)—;

qb=0-3

nb=1-3

mb=1-3

pb=0-3

R^(b3) is selected from H, R^(b1)—, R^(b2)—C(O)—, or R^(b2)—S(O₂)—;

R^(b4), R^(b5) and R^(b6) are, independently, selected from a groupconsisting of H, or C₁-C₄ alkyl, and

Xaa₁ is selected from the group consisting of a direct bond, aconventional α-amino acid; a non-conventional α-amino acid; a β-aminoacid; a γ-amino acid; or a residue of Formula IIa-y:

R^(1a) is selected from H, C₁-C₆ alkyl;

R^(1b) is selected from H, C₁-C₆ alkyl optionally substituted by OH,hydroxyC₁-C₆ alkyl optionally substituted by OH;

R^(1c) is selected from H, C₁-C₆ alkyl;

R^(1d) is selected from H, C₁-C₆ alkyl;

R^(1a) and R^(1b) together may form a heterocyclic ring;

n¹ is 0 to 3;

Xaa₂ is an amino acid residue of Formula IIIa-g:

wherein

R^(2a) is selected from the group consisting of H, methyl, ethyl,propyl, isopropyl, C₁-C₂ alkyl C₃-C₇ cycloalkyl and aryl C₁-C₂ alkyl;

R^(2b) and R^(2c) are, independently, selected from the group consistingof H, methyl, ethyl, propyl; and isopropyl, with the proviso that atleast one of R^(2b) and R^(2c) is H;

R^(2d) represents from 0 to 3 substituents, each such substituent being,independently, selected from the group consisting of H, Cl, F, Br, NO₂,NH₂, CN, CF₃, OH, OR^(2e) and C₁-C₄ alkyl;

R^(2a) and R^(2b) or R^(2a) and R^(2c) together may form a heterocyclicring;

R^(2e) is selected from the group consisting of methyl, ethyl, propyl,and isopropyl; or

Xaa₁ and Xaa₂ together may be selected from an amino acid residue ofFormula IVa-b

Xaa₃ is selected from the group consisting of Gly, Ala, a conventionalD-α-amino acid, a non-conventional D-α-amino acid, and an amino acidresidue of Formula Va:

wherein R^(3a) is selected from the group consisting of H or C₁-C₄alkyl;

R^(3b) is selected from the group consisting of H, —(CH₂)_(n3a)—X^(3a);

n3a is 1 to 5;

X^(3a) is selected from the group consisting of H, NR^(3c)R^(3d);

R^(3c) and R^(3d) are independently selected from a group consisting ofH, C₁-C₈ alkyl, —(C═N)—NH₂ and —(CH₂)_(n3b)X^(3b);

n3b is 1 to 4;

X^(3b) is selected from the group consisting of NR^(3e)R^(3f), C₅-C₆heteroaryl, C₄-C₇ heterocyclyl, —NHC(═N)NH₂;

R^(3a) and R^(3f) are independently selected from a group consisting ofH, C₁-C₈ alkyl,

wherein R^(3e) and R^(3f) can form a cyclic structure;

R^(3a) and R^(3b) can be linked to form a cyclic structure;

or R^(3a) and R^(3b) can be linked with a heteroatom selected from thegroup consisting of N, O, and S, to form a heterocyclic structure;

or

Xaa₂ and Xaa₃ together may be selected from an amino acid residue ofFormula Vb:

wherein R^(3g) represents from 0 to 3 substituents, each suchsubstituent being, independently, selected from the group consisting ofH, Cl, F, Br, NO₂, NH₂, CN; CF₃, OH, OR^(3h) and C₁-C₄ alkyl;

R^(3h) is selected from the group consisting of C₁-C₄ alkyl

Xaa₄ is an amino acid residue of Formula VIa-h:

wherein R^(4a) is selected from the group consisting of H, C₁-C₈ alkylwhich may be substituted with a moiety selected from the groupconsisting of OH, CO₂R^(4c), C(═O)—NH₂, a 5-6 membered heteroaryl,C₁-C₁₀ alkyl, C₅-C₈ cycloalkyl C₁-C₁₀ alkyl, and C₅-C₈ cycloalkyl,—(CH₂)_(n4a)—X^(4a);

n^(4a) is 1 or 2;

R^(4b) is selected from the group consisting of H and methyl;

R^(4c) is selected from the group consisting of H, and C₁-C₃alkyl; and

X^(4a) is OH, CO₂R^(4d), NR^(4e)R^(4f), SR^(4g), 4-hydroxyphenyl;

R^(4d), R^(4e) and R^(4f) independently are selected from the groupconsisting of H, and C₁-C₃ alkyl;

R^(4g) is selected from the group consisting of C₁-C₃ alkyl;

m4a, and m4b are independently selected from 0 or 1;

R^(4h) is C₂-C₆ alkyl;

or

Xaa₃ and Xaa₄ together may be selected from an amino acid residue ofFormula VIb-h;

Xaa₅ is an amino acid residue of Formula VII:

wherein R^(5a) is (CH₂)_(n5a)—X^(5a);

n5a is 1 to 6;

X^(5a) is selected from the group consisting of H, NH₂, and a C₄₋₇amine-containing aliphatic heterocyclic ring;

R^(5b) is selected from the group consisting of H and methyl;

R^(5c) is selected from the group consisting of H and methyl;

and wherein R^(5c) and R^(5a) can combine to form a four to six memberedheterocyclic ring or can be linked with a heteroatom selected from thegroup consisting of N, O, and S to form a monocyclic or bicyclicheterocyclic structure; wherein said heterocyclic ring may have from 0to 3 substituents, each such substituent being, independently, selectedfrom from the group consisting of OH, OR^(5d), F, C₁-C₄ alkyl,—NHC(═NH)NH₂, aryl and NR^(5e)R^(5f);

R^(5d) is selected from C₁-C₄ alkyl, C₁-C₄ alkylaryl;

R^(5e) is selected from the group consisting of H, C₁-C₄ alkyl,—C(═O)(CH₂)_(n5b)—X^(5b), —CH₂(CH₂)_(n5c)—X^(5b);

R^(5f) is selected from the group consisting of H, C₁-C₄ alkyl,—CH₂(CH₂)_(n5d)—X^(5c);

n5b is selected from the group consisting of 1, 2, 3, and 4;

n5c and n5d are independently selected from the group consisting of 2,3, and 4;

X^(5b) and X^(5c) are independently selected from the group consistingof H, NR^(5g)R^(5h);

R^(5g) and R^(5h) are independently selected from a group consisting ofH, C₁-C₄ alkyl;

Xaa₆ is an amino acid residue of Formula VIIIa-d:

wherein R^(6a) is selected from the group consisting of C₁-C₈ alkyl,aryl C₁-C₄ alkyl, C₄-C₇ cycloalkyl C₁-C₄ alkyl, C₁-C₄ alkylS(C₁-C₄alkyl), and C₄-C₇ cycloalkyl, wherein said C₁-C₈ alkyl and C₄-C₇cycloalkyl may be substituted with a moiety selected from the groupconsisting of OH, O(C₁-C₄ alkyl), S(C₁-C₄ alkyl), and NR^(6d)R^(6e);

R^(6b) is H;

R^(6c) is selected from the group consisting of H, and C₁-C₄alkyl;

R^(6d), and R^(6e) are, independently, selected from the groupconsisting of H, and C₁-C₄ alkyl;

wherein R^(6a) and R^(6c) can form a cyclic structure, which may besubstituted with a moiety selected from the group consisting of OH,C₁-C₄ alkyl, NH₂ and F;

or R^(6a) and R^(6c) can be linked with a heteroatom selected from thegroup consisting of N, O, and S, to form a heterocyclic structure;

or

Xaa₅ and Xaa₆ together may be an amino acid residue of Formula VIIIe:

Xaa ₇ is an amino acid residue of Formula IXa-b:

wherein R^(7a) is selected from the group consisting of C₁-C₄ alkyl,C₃-C₇ cycloalkyl, 2-thienyl, (CH₂)_(n7a)—X^(7a), and C₁-C₄ alkylsubstituted with OH;

R^(7b) is H, and 2-thienyl;

R^(7c) is selected from a group consisting of H, and methyl;

R^(7d) is C₁-C₄ alkyl;

n^(7a) is selected from the group consisting of 1 and 2;

X^(7a) is selected from the group consisting of 2-thienyl, C(═O)OR^(7e),C(═O)NH₂, S(═O)₂OH, OS(═O)₂OH, B(OH)₂, P(═O)(OH)₂, and OP(═O)(OH)₂;

wherein R^(7e) is selected from the group consisting of H, and C₁-C₄alkyl;

Xaa₈ is an amino acid residue of Formula Xa-g:

wherein R^(8a) is selected from the group consisting of(CH₂)_(m8a)—X^(8a), and a C₄-C₇ nitrogen-containing aliphaticheterocyclic ring;

m^(8a)=1-5;

X^(8a) is selected from the group consisting of H, NH₂, and—NHC(═NH)NH₂;

R^(8b) is selected from the group consisting of H and methyl;

R^(8c) is selected from the group consisting of H, NH₂, and OH;

Y^(8a) is selected from the group consisting of CH(R^(8d)), and S;

R^(8d) is selected from the group consisting H, aryl, and OH;

Y^(8b) is selected from the group consisting of CH(R^(8e)), and NH;

R^(8e) is selected from the group consisting H, NH₂ and OH;

Y^(8c) is selected from the group CH₂, and NR^(8f);

R^(8f) is selected from the group H, —C(═NH)NH₂, and —C(═O)CH₂NH₂;

or

Xaa₇ and Xaa₈ together may be an amino acid residue of Formula Xh:

Xaa₉ is selected from the group consisting of a direct bond, and anamino acid residue of Formula XIa-c,

wherein R^(9a) is selected from the group consisting of C₁-C₅ alkyl, andC₄-C₇ cycloalkyl;

R^(9b) is selected from the group consisting of H, C₁-C₅ alkyl;

and wherein R^(9a) and R^(9b) can form a 5-7 membered cycloalkyl ring;

R^(9a) is selected from the group consisting of H, methyl;

or

Xaa₈ and Xaa₉ together may be a residue of Formula XId:

and

Z is selected from the group consisting of H, OR^(11a), NHR^(11b) aconventional α-amino acid, a non-conventional α-amino acid, a β-aminoacid; and a peptide consisting of from 2 to 30 amino acids selected fromthe group consisting of conventional α-amino acids, non-conventionalα-amino acids, and β-amino acids;

wherein R^(11a) and R^(11b) are independently selected from the groupconsisting of H, C₁-C₈ alkyl, C₄-C₈ cycloalkyl, C₇-C₁₂ bicycloalkyl,C₇-C₁₂ cycloalkylaryl, C₁-C₄ alkyl C₄-C₈ cycloalkyl, or a residue offormula XIIa-c:

As used herein, the phrase “optionally substituted” shall be understoodby the skilled artisan to mean that the moiety to which the phraserefers may be unsubstituted, or it may be substituted with certainspecified additional moieties. For example, the phrase “C₁-C₁₂ alkyloptionally substituted by NR^(b4)R^(b5) OH, OR^(b6), C₃-C₈ cyclic alkyl,aryl, heteroaryl, or heterocyclyl” refers to a C₁-C₁₂ alkyl compoundthat is either non-substituted or is substituted by a moiety selectedfrom the group consisting of NR^(b4)R^(b5), OH, OR^(b6), C₃-C₈ cyclicalkyl, aryl, heteroaryl, and heterocyclyl. The compound, hexane, wouldbe considered a C₆ alkyl compound that is not substituted, while thecompound 3-hexanol is a C₆ alkyl compound that is substituted on thethird carbon atom with an OH moiety.

In certain preferred NPR-B agonists of the invention:

B is selected from the group consisting of R^(b1)—, R^(b2)—C(O)—;

R^(b1) is selected from C₁-C₁₂ alkyl optionally substituted byNR^(b4)R^(b5);

R^(b2) is selected from C₁-C₁₂ alkyl optionally substituted byNR^(b4)R^(b5);

R^(b4), and R^(b5) are, independently, selected from a group consistingof H, and C₁-C₄ alkyl, and

Xaa₁ is selected from the group consisting of a direct bond, aconventional α-amino acid; a non-conventional α-amino acid; a β-aminoacid; or a residue selected from the group consisting of Formula IIa,IIs, IIt, IIu, and IIv:

R^(1a) is selected from H, C₁-C₆ alkyl;

R^(1b) is selected from H, C₁-C₆ alkyl optionally substituted by OH,hydroxyC₁-C₆ alkyl optionally substituted by OH;

R^(1c) is selected from H, C₁-C₆ alkyl;

R^(1a) and R^(1b) together may form a heterocyclic ring;

n¹ is 0 to 3; and

Xaa₂ is an amino acid residue of Formula IIIc or Formula IIIb:

wherein

R^(2a) is selected from the group consisting of H, methyl, ethyl,propyl, isopropyl, C₁-C₂ alkyl C₃-C₇ cycloalkyl and aryl C₁-C₂ alkyl;

R^(2b) and R^(2c) are, independently, selected from the group consistingof H, methyl, ethyl, propyl; and isopropyl, with the proviso that atleast one of R^(2b) and R^(2c) is H;

R^(2d) represents from 0 to 3 substituents, each such substituent being,independently, selected from the group consisting of H, Cl, F, Br, NO₂,NH₂, CN, CF₃, OH, OR^(2e) and C₁-C₄ alkyl;

R^(2a) and R^(2b) or R^(2a) and R^(2c) together may form a heterocyclicring;

R^(2e) is selected from the group consisting of methyl, ethyl, propyl,and isopropyl; and

Xaa₃ is an amino acid residue of Formula Va:

wherein R^(3a) is selected from the group consisting of H or C₁-C₄alkyl;

R^(3b) is selected from the group consisting of H, —(CH₂)_(n3a)—X^(3a);

n3a is 1 to 5;

X^(3a) is selected from the group consisting of H, NR^(3c)R^(3d);

R^(3c) and R^(3d) are independently selected from a group consisting ofH, C₁-C₈ alkyl, —(C═N)—NH₂ and —(CH₂)_(n3b)X^(3b),

n3b is 1 to 4;

X^(3b) is selected from the group consisting of NR^(3e)R^(3f), C₅-C₆heteroaryl, C₄-C₇ heterocyclyl, —NHC(═N)NH₂;

R^(3e) and R^(3f) are independently selected from a group consisting ofH, C₁-C₈ alkyl,

wherein R^(3e) and R^(3f) can form a cyclic structure;

R^(3a) and R^(3b) can be linked to form a cyclic structure; or R^(3a)and R^(3b) can be linked with a heteroatom selected from the groupconsisting of N, O, and S, to form a heterocyclic structure;

and

Xaa is an amino acid residue of Formula VIa:

wherein R^(4a) is selected from the group consisting of H, C₁-C₈ alkylwhich may be substituted with a moiety selected from the groupconsisting of OH, CO₂R^(4c), C(═O)—NH₂, a 5-6 membered heteroaryl,—C₁-C₁₀ alkyl, C₅-C₈ cycloalkyl C₁-C₁₀ alkyl, and C₅-C₈ cycloalkyl;

n4a is 1 or 2;

R^(4b) is selected from the group consisting of H and methyl;

R^(4a) is selected from the group consisting of H, and C₁₋₃alkyl; and

and

Xaa₅ is an amino acid residue of Formula VII:

wherein R^(5a) is (CH₂)_(n5a)—X^(5a);

n5a is 1 to 6;

X^(5a) is selected from the group consisting of H, NH₂, and a C₄₋₇amine-containing aliphatic heterocyclic ring;

R^(5b) is selected from the group consisting of H and methyl;

R^(5c) is selected from the group consisting of H and methyl;

and wherein R⁵ and R^(5a) can combine to form a four to six memberedheterocyclic ring wherein said heterocyclic ring may have from 0 to 2substituents, each such substituent being, independently, selected fromfrom the group consisting of OH, OR^(5d), F, C₁-C₄ alkyl, —NHC(═NH)NH₂,aryl and NR^(5e)R^(5f);

R^(5d) is selected from C₁-C₄ alkyl, C₁-C₄ alkylaryl;

R^(5c) is selected from the group consisting of H, C₁-C₄ alkyl,—C(═O)(CH₂)_(n5b)—X5b, —CH₂(CH₂)_(n5c)—X^(5b);

R^(5f) is selected from the group consisting of H, C₁-C₄ alkyl,—CH₂(CH₂)_(n5d)—X^(5c);

n5b is selected from the group consisting of 1, 2, 3, and 4;

n5c and n5d are independently selected from the group consisting of 2,3, and 4;

X^(5b) and X^(5c) are independently selected from the group consistingof H, NR^(5g)R^(5h);

R^(5g) and R^(5h) are independently selected from a group consisting ofH, C₁-C₄ alkyl and

Xaa₆ is an amino acid residue of Formula VIIIa:

wherein R^(6a) is selected from the group consisting of C₁-C₈ alkyl,aryl C₁-C₄ alkyl , C₄-C₇ cycloalkyl C₁-C₄ alkyl, C₁-C₄ alkylS(C₁-C₄alkyl), and C₄-C₇cycloalkyl, wherein said C₁-C₈ alkyl and C₄-C₇cycloalkyl may be substituted with a moiety selected from the groupconsisting of OH, O(C₁-C₄ alkyl), and S(C₁-C₄ alkyl);

R^(6b) is H;

R^(6c) is selected from the group consisting of H, and C₁-C₄alkyl; and

Xaa₇ is an amino acid residue of Formula IXa:

wherein R^(7a) is selected from the group consisting of C₁-C₄ alkyl,C₃-C₇ cycloalkyl, 2-thienyl, and C₁-C₄ alkyl substituted with OH;

R^(7b) is H, and 2-thienyl;

R^(7c) is selected from a group consisting of H, and methyl;

and

Xaa₈ is an amino acid residue of Formula X(a)-(g):

wherein R^(8a) is (CH₂)_(n8a)—X^(8a);

m^(8a)=1-5;

X^(8a) is selected from the group consisting of H, NH₂, and—NHC(═NH)NH₂;

R^(8b) is selected from the group consisting of H and methyl; and

Xaa₉ is selected from the group consisting of a direct bond, and anamino acid residue of Formula XIa-c,

wherein R^(9a) is selected from the group consisting of C₁-C₅ alkyl, andC₄-C₇ cycloalkyl;

R^(9b) is selected from the group consisting of H, and C₁-C₅ alkyl;

or R^(9a) and R^(9b) can form a 5-7 membered cycloalkyl ring;

R^(9c) is selected from the group consisting of H, and methyl;

and

Z is NHR^(11b);

wherein R^(11b) is selected from the group consisting of H, C₁-C₈ alkyl,C₄-C₈ cycloalkyl, C₇-C₁₂ bicycloalkyl, C₇-C₁₂ cycloalkylaryl, C₁-C₄alkyl C₁-C₈ cycloalkyl, or a residue of formula XIIa-c

In more preferred embodiments of the present invention, B is selectedfrom the group consisting of R^(b1)—, and R^(b2)—C(O)—;

R^(b1) is selected from the group consisting of C₆-C₁₀ alkyl and C₆-C₁₀alkyl substituted by NR^(b4)R^(b5);

R^(b2) is selected from the group consisting of C₆-C₁₀ alkyl and C₆-C₁₀alkyl substituted by NR^(b4)R^(b5);

R^(b4), and R^(b5) are, independently, selected from a group consistingof H, and C₁-C₄ alkyl, and

Xaa₁ is selected from the group consisting of a direct bond, aconventional α-amino acid; a non-conventional α-amino acid; a β-aminoacid; a residue of Formula IIa, a residue of Formula IIs, a residue ofFormula IIt, a residue of Formula IIu, and a residue of Formula IIv

wherein R^(1a) is selected from H, and C₁-C₄ alkyl;

R^(1b) is selected from H, C₁-C₄ alkyl optionally substituted by OH, andhydroxy C₁-C₄ alkyl optionally substituted by OH;

R^(1c) is selected from H, C₁-C₆ alkyl;

R^(1a) and R^(1b) together may form a heterocyclic ring;

n¹ is 0, 1; and

Xaa₂ is an amino acid residue of Formula III:

wherein

R^(2a) is selected from the group consisting of H, methyl, ethyl,propyl, isopropyl, C₁-C₂ alkyl C₃-C₇ cycloalkyl and awl C₁-C₂ alkyl;

R^(2b) and R^(2c) are, independently, selected from the group consistingof H, methyl, ethyl, propyl; and isopropyl, with the proviso that atleast one of R^(2b) and R^(2c) is H;

R^(2d) represents from 0 to 3 substituents, each such substituent being,independently, selected from the group consisting of H, Cl, F, Br, CN,CF₃, OH, OR^(2e) and C₁-C₄ alkyl;

R^(2e) is selected from the group consisting of methyl, ethyl, propyl,and isopropyl; and

Xaa₃ is an amino acid residue of Formula Va:

wherein R^(3a) is selected from the group consisting of H and C₁-C₄alkyl;

R^(3b) is selected from the group consisting of H, and—(CH₂)_(n3a)—X^(3a);

n3a is 1 to 5;

X^(3a) is selected from the group consisting of H, and NR^(3c)R^(3d);

R^(3a) and R^(3d) are independently selected from a group consisting ofH, C₁-C₈ alkyl, and —(C═N)—NH₂;

R^(3a) and R^(3b) can be linked to form a cyclic structure;

or R^(3a) and R^(3b) can be linked with a heteroatom selected from thegroup consisting of N, O, and S, to form a heterocyclic structure;

and

Xaa₄ is an amino acid residue of Formula VIa:

wherein R^(4a) is selected from the group consisting of H, C₁-C₈ alkylwhich may be substituted with a moiety selected from the groupconsisting of OH, and CO₂R^(4c);

R^(4b) is selected from the group consisting of H and methyl;

R^(4c) is selected from the group consisting of H, and C₁-C₃alkyl; and

and

Xaa₅ is an amino acid residue of Formula VII:

wherein R^(5a) is (CH₂)_(n5a)—X^(5a);

n5a is 1 to 6;

X^(5a) is selected from the group consisting of H, NH₂, and a C₄₋₇amine-containing aliphatic heterocyclic ring;

R^(5b) is selected from the group consisting of H and methyl;

R^(5c) is selected from the group consisting of H and methyl;

and wherein R^(5c) and R^(5a) can combine to form a four to six memberedheterocyclic ring wherein said heterocyclic ring may have from 0 to 2substituents, each such substituent being independently selected fromfrom the group consisting of OH, F, C₁-C₄ alkyl, —NHC(═NH)NH₂, aryl andNR^(5e)R^(5f);

R^(5f) is selected from the group consisting of H, C₁-C₄ alkyl,—C(═O)(CH₂)_(n5b)—X^(5b), and —CH₂(CH₂)_(n5c)—X^(5b);

R^(5f) is selected from the group consisting of H, C₁-C₄ alkyl, and—CH₂(CH₂)_(n5d)—X^(5c);

n5b is selected from the group consisting of 1, 2, 3, and 4;

n5c and n5d are independently selected from the group consisting of 2,3, and 4;

X^(5b) and X^(5a) are independently selected from the group consistingof H, and NR^(5g)R^(5h);

R^(5g) and R^(5h) are independently selected from a group consisting ofH, and C₁-C₄ alkyl and

Xaa₆ is an amino acid residue of Formula VIIIa:

wherein R^(6a) is selected from the group consisting of C₁-C₈ alkyl,aryl C₁-C₄ alkyl , C₄-C₇ cycloalkyl C₁-C₄ alkyl, and C₄-C₇cycloalkyl,wherein said C₁-C₈ alkyl and C₄-C₇ cycloalkyl may be substituted with amoiety selected from the group consisting of OH, and O(C₁-C₄ alkyl);

R^(6b) is H;

R^(6c) is selected from the group consisting of H, and C₁-C₄alkyl; and

Xaa₇ is an amino acid residue of Formula IX:

wherein R^(7a) is selected from the group consisting of C₁-C₄ alkyl,C₃-C₇ cycloalkyl, 2-thienyl, and C₁-C₄ alkyl substituted with OH;

R^(7b) is H, and 2-thienyl;

R^(7c) is selected from a group consisting of H, and methyl;

and

Xaa₈ is an amino acid residue of Formula Xa:

wherein R^(8a) is (CH₂)_(m8a)—X^(8a);

m^(8a)=1-5;

X^(8a) is selected from the group consisting of H, NH₂, and—NHC(═NH)NH₂;

R^(8b) is selected from the group consisting of H and methyl; and

Xaa₉ is selected from the group consisting of a direct bond, and anamino acid residue of Formula XIa,

wherein R^(9a) is selected from the group consisting of C₁-C₅ alkyl, andC₄-C₇ cycloalkyl;

R^(9b) is selected from the group consisting of H, and C₁-C₅ alkyl;

and wherein R^(9a) and R^(9b) can form a 5-7 membered cycloalkyl ring;

R^(9c) is selected from the group consisting of H, and methyl;

and

Z is NHR^(11b);

wherein R^(11b) is selected from the group consisting of H, C₁-C₈ alkyl,C₄-C₈ cycloalkyl, C₇-C₁₂ bicycloalkyl, C₇-C₁₂ cycloalkylaryl, and C₁-C₄alkyl C₄-C₈ cycloalkyl.

The sequences of the preferred novel NPR-B agonists of the invention areprovided herein in typical peptide sequence format, as would beunderstood by the ordinary skilled artisan. For example, thethree-letter code of a conventional amino acid, or the abbreviation fora non-conventional amino acid, indicates the presence of a particularamino acid in a specified position in the sequence of the molecule, eachamino acid being connected to the next and/or previous amino acid by ahyphen. The hyphen, which represents a chemical bond, typically an amidebond, removes OH from the 1-carboxyl group of the amino acid when it isplaced right of the abbreviation, and removes H from the 2-amino group(or the only present amino group in case of amino acids lacking a2-amino group, e.g., Bal) of the amino acid when it is placed on theleft of the abbreviation. It is understood that both modifications canapply to one amino acid.

In the case of additional functional groups in the side chains ofconventional or non-conventional amino acids, only the 2-amino and/orthe 1-carboxy group is used for the formation of peptide bonds.

The C-termini of the novel NPR-B agonists described herein are shown inexplicit form by adding either OH, NH2 or an abbreviation for a specificterminating amine separated by a hyphen on the right of the abbreviationof the C-terminal amino acid.

These specific terminating amines are provided in Table 2 as fullformulas and similar conventions with regard to hyphens and itsstructure in a peptide context apply to them, e.g.,

3791=NH₂—CH(CH₂—CH₃)—CH₂—CH₃

−3791=—NH—CH(CH₂—CH₃)—CH₂—CH₃

The N-termini of the novel peptides described herein are shown inexplicit form by adding either H (for a free N-terminus), or anabbreviation for a specific terminating carboxylic acid, sulfonic acidor another terminating group in front of the symbol of the N-terminalamino acid.

These specific terminating carboxylic acids, sulfonic acids or otherterminating groups like alkyl are provided in Table 2 as full formulasand similar conventions with regard to hyphens and its structure in apeptide context apply to them, e.g.,

Hex=−Hexanoic acid

Hex−=Hexanoyl−.

For conventional amino acids and some non-conventional amino acids, a3-letter code was used where the first letter indicates thestereochemistry of the C-alpha-atom. For example, a capital first letterindicates that the L-form of the amino acid is present in the peptidesequence, while a lower case first letter indicates that the D-form ofthe correspondent amino acid is present in the peptide sequence.

In preferred embodiments of the present invention, the novel NPR-Bagonist is an 8-13 amino acid peptide having a sequence as set forth inTable 3. The agonistic activity of the preferred compounds is alsoprovided in Table 3 and was categorized based upon the followingconventions:

NPR-B activation (assayed with GTM-3 Cells) EC₅₀ Emax (CNP = 100%) Group≦1 μM >50% A ≦5 μM >20% B ≦15 μM  >10% C

The agonistic activity data of each compound was checked first todetermine whether it fulfills the criteria for the activity group A. Ifit did not fulfill the criteria for activity group A, it was checked forgroup B criteria. If it did not fulfill the criteria for activity groupA or activity group B, it was finally checked for group C criteria. Ifit did not fulfill the criteria for activity group C, it was notincluded in Table 3.

All examples in Table 3 are linear peptides written in three letter codewhere applicable. For non-conventional amino acids and other chemicalmoieties the abbreviations which are listed in Table 2 were used. Invitro activities reported in Table 3 resulted from experiments performedaccording to the methods described in Example 4.

In certain embodiments of the NPR-B agonists of the invention, in thecompound of Formula 1:

B will be selected from a bond, Occ, Oct, Sbt, 1319, 1320, and 5587;

Xaa₁ will be selected from Gly, AR-201-49, AR-201-68, ala, abu, his,aze, pro, pip, thz, thi, asn, ser, His, Ala, Ser, Bal, Sni, Az3, andGab;

Xaa₂ will be selected from Phe, Pcf, Nmf, Pbf, Pff, Pmf, Eaa. Mcf, Thk,and Mtf;

Xaa₃ will be selected from Gly, Aib, Ebc, a conventional D-α-amino acid,and a non-conventional D-α-amino acid, and will preferably be selectedfrom Gly, Fhy, Apc, Egz, Aib, Ebc, ala, lys, lys(Me2), arg, leu, nle,ctb, abu, AR-385-12, Egg, ser, orn, orn(Me2), and dap(Me₂);

Xaa₄ will be selected from Leu, Nva, Nle, Hle, Npg, Cha, and Ala;

Xaa₅ will be selected from Lys, Orn, Hly, Hpa, Dab, Arg, N(alkyl)derivatives of any of the preceding amino acids, Nmk, Hpr, Pro, Tfp,Apr, Eaz, Hyp, Tap, Tap(G), Tap(Bal), Tap(Et), Tap(Ae), Tap(Ap), Amp,Pip, and Chy;

Xaa₆ will be selected from a bond, Leu, Ile, Nml, Tap, Npg, SH-158,Dap(Me2), Cpg, Val, Tbg, Chg, Hle, Nle, and N(alkyl) derivatives of anyof the preceding amino acids;

Xaa₇ will be selected from Asp, Val, BB725, BB727, Ser, Thr, and Cya;

Xaa₈ will be selected from Arg, Nmr, Pro, Eaz, Pca, Orn, Fhz, Har, Nar,Cyr, Mmr, Dmr, Bmr, Opy, and N(alkyl) derivatives of any of thepreceding amino acids;

Xaa₉ will be selected from Ile, Tbg, Deg, Egz, Aml, 1860, Che, Nmi, Leu,Val, Ecb, and Eca; and

Xaa₁₀ will be selected from a bond, Ser and a N(alkyl) derivativethereof.

TABLE 3 Preferred compounds according to the present inventionand their agonistic activity in in vitro assays. SEQ ID (M + H)⁺ inActivity Structure JAL NO: MS [amu] (group)Hex-Ebe-pro-Phe-Gly-Leu-Pro-Ile-Asp-Arg-Ile- JAL- 19 1446 C Ser-Ebe-NH₂;0533 Hex-Ebe-pro-Phe-Gly-Leu-Lys-Ile-Asp-Arg-Ile- JAL- 20 1477 CSer-Ebe-NH₂; 0534 Hex-Ser-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 211391 C Ser-Ser-NH₂; 0535 Hex-Ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-JAL- 22 1359 B Ser-Ala-NH₂; 0536Hex-Ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 23 1345 C Ser-Gly-NH₂;0537 Hex-Gly-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 24 1345 BSer-Ala-NH₂; 0538 Hex-Gly-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 251331 B Ser-Gly-NH₂; 0539 Hex-Ebe-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-JAL- 26 1334 C Ser-NH₂; 0540Hex-Ebe-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 27 1247 C NH₂; 0541Hex-Gab-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-Ser- JAL- 28 1348 C Ebe-NH₂;0542 Hex-Mam-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-Ser- JAL- 29 1396 CEbe-NH₂; 0543 Hex-Gly-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 30 1188C NH₂; 0631 Hex-Ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 31 1202 CNH₂; 0632 Hex-Ser-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 32 1218 CNH₂; 0633 Hex-Pro-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 33 1228 CNH₂; 0634 Hex-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 34 1201 CNH₂; 0635 Hex-Gly-pro-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 35 1213 CNH₂; 0636 Hex-Ser-pro-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 36 1241 CNH₂; 0638 Hex-Mam-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 37 1193 C NH₂;0647 Hex-Pam-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 38 1193 C NH₂; 0648Hex-Mpe-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 39 1193 C NH₂; 0649Hex-Ppe-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 40 1193 C 0650Hex-Inp-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 41 1171 C 0651Hex-Acp-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 42 1210 C NH₂; 0652Hex-Fir-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 43 1199 C 0653Hex-Nip-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 44 1171 C 0654Hex-Eah-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 45 1228 C NH₂; 0656Hex-Fio-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 46 1185 C 0657Hex-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Eca- JAL- 47 1199 C NH₂; 06921339-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 48 1255 C NH₂; 0693Occ-pro-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 49 1184 C NH₂; 06941339-pro-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 50 1210 C NH₂; 06951320-pro-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 51 1218 C NH₂; 0696Occ-Nip-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 52 1198 B 0697Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 53 1229 B NH₂; 07011340-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 54 1241 C NH₂; 0703Hex-Tnc-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 55 1186 C NH₂; 0713Hex-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Chg- JAL- 56 1227 C NH₂; 0718Hex-ala-ala-Phe-Paa-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 57 1157 C 0731Occ-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 58 1158 C 0738Occ-thz-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 59 1202 C 0739Occ-aze-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 60 1170 C 0740Occ-Az3-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 61 1170 C NH₂; 0742Occ-Sni-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 62 1198 B 0743Occ-Rni-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 63 1198 C 0744Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-2137; JAL- 64 1199 C 0748Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-3816; JAL- 65 1201 C 0749Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-3806; JAL- 66 1187 C 0751Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-565; JAL- 67 1200 B 0752Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-2797; JAL- 68 1252 B 0754Occ-val-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 69 1186 C 0756Occ-tbg-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 70 1200 C 0758Occ-Amcp-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile- JAL- 71 1184 C NH₂; 0760Occ-Ebc-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 72 1170 C 0761Occ-abu-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 73 1171 C 0762Occ-ser-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 74 1174 C 0763Occ-ala-ala-Phe-Gly-Leu-Lys-leu-Asp-Arg-Ile- JAL- 75 1229 C NH₂; 0769Occ-ala-ala-Phe-Gly-Leu-Lys-Ile-Asp-Arg-Ile- JAL- 76 1229 C NH₂; 0770Occ-ala-ala-Phe-Gly-Leu-Lys-Val-Asp-Arg-Ile- JAL- 77 1215 C NH₂; 0771Occ-ala-ala-Phe-Gly-Leu-Lys-Chg-Asp-Arg-Ile- JAL- 78 1255 C NH₂; 0772Occ-ala-ala-Phe-Gly-Leu-Lys-Nle-Asp-Arg-Ile- JAL- 79 1229 C NH₂; 0775Occ-ala-ala-Phe-Gly-Leu-Lys-Nml-Asp-Arg-Ile- JAL- 80 1243 C NH₂; 0776Occ-ala-ala-Phe-Gly-Leu-Pro-Leu-Asp-Arg-Ile- JAL- 81 1214 B NH₂; 0781_01Occ-ala-ala-Phe-Gly-Leu-Nmk-Leu-Asp-Arg-Ile- JAL- 82 1243 C NH₂; 0782933-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 83 1208 C 07861270-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 84 1160 C 07874956-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Ile-NH₂; JAL- 85 1144 C 0788Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-1860; JAL- 86 1213 B 0789Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-504; JAL- 87 1251 C 0790Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-559; JAL- 88 1185 C 0791Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-3791; JAL- 89 1187 C 0792Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Che; JAL- 90 1212 B 0797Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-1859; JAL- 91 1211 C 0798Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-1934; JAL- 92 1304 B 0799Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-1906; JAL- 93 1209 B 0801Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-873; JAL- 94 1192 C 0824Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-5116; JAL- 95 1241 C 0825Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-5119; JAL- 96 1270 B 0826Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-5118; JAL- 97 1270 C 0831Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-5163; JAL- 98 1227 C 0833Occ-ala-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-5164; JAL- 99 1255 C 0834Occ-ala-Phe-Gly-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 100 1127 C 0835Occ-pro-Phe-Gly-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 101 1153 C 0836Occ-Sni-Phe-Gly-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 102 1167 C 0837Occ-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-1860; JAL- 103 1141 B 0839Occ-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-Che; JAL- 104 1141 C 0840Occ-ala-Phe-Gly-Leu-Lys-Leu-Asp-Arg-5121; JAL- 105 1143 C 0841Occ-ala-Phe-Gly-Leu-Pro-Ile-Asp-Arg-Ile-NH₂; JAL- 106 1127 C 0894Occ-ala-Phe-Gly-Leu-Pro-Nml-Asp-Arg-Ile-NH₂; JAL- 107 1141 B 0895Occ-ala-Phe-Gly-Leu-Pro-Npg-Asp-Arg-Ile-NH₂; JAL- 108 1141 C 0896Occ-ala-Phe-Gly-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 109 1143 B 0898Occ-ala-Phe-Gly-Npg-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 110 1141 C 0903Occ-ala-Nmf-Gly-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 111 1141 C 0906Occ-ala-Phe-Gly-Leu-Pro-Leu-Asp-Nmr-Ile-NH₂; JAL- 112 1141 C 0921Occ-ala-Phe-Gly-Leu-Pro-Leu-Asn-Arg-Ile-NH₂; JAL- 113 1127 C 0924Occ-ala-Phe-Gly-Leu-Pro-Leu-Nva-Arg-Ile-NH₂; JAL- 114 1111 C 0926Occ-ala-Phe-Gly-Leu-Pro-Leu-Val-Arg-Ile-NH₂; JAL- 115 1111 C 0927Occ-ala-Phe-Gly-Leu-Pro-Leu-Thr-Arg-Ile-NH₂; JAL- 116 1113 C 0929Occ-ala-Phe-Gly-Cha-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 117 1167 C 0940Occ-ala-Phe-Gly-Nle-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 118 1127 C 0942Occ-ala-Phe-Aib-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 119 1155 C 0943Occ-ala-Phe-ala-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 120 1141 C 0944Occ-ala-Phe-Ebc-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 121 1153 C 0945Occ-ala-Mcf-Gly-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 122 1161 C 0946Occ-Sar-Phe-Gly-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 123 1127 C 0950Occ-Gly-Phe-Gly-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 124 1113 C 0951Occ-aze-Phe-Gly-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 125 1139 B 0953Occ-ala-Nmf-Gly-Leu-Pro-Nml-Asp-Arg-Ile- JAL- 126 1155 B NH₂; 0954Occ-pro-Phe-Gly-Leu-Pro-Nml-Asp-Arg-Ile-NH₂; JAL- 127 1167 B 0955_01Occ-Sni-Phe-Gly-Leu-Pro-Nml-Asp-Arg-Ile-NH₂; JAL- 128 1181 B 0956Occ-pro-Nmf-Gly-Leu-Pro-Nml-Asp-Arg-Ile- JAL- 129 1181 C NH₂; 0957Occ-Sni-Nmf-Gly-Leu-Pro-Nml-Asp-Arg-Ile- JAL- 130 1195 B NH₂; 0958_01Occ-ala-Phe-Gly-Leu-Pro-Hle-Asp-Arg-Ile-NH₂; JAL- 131 1141 C 0959Occ-ala-Phe-Gly-Leu-Amp-Leu-Asp-Arg-Ile- JAL- 132 1141 C NH₂; 0962Occ-ala-Phe-Gly-Leu-Chy-Leu-Asp-Arg-Ile-NH₂; JAL- 133 1143 C 0964Occ-pro-Nmf-Gly-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 134 1167 C 0966Occ-Sni-Nmf-Gly-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 135 1181 C 0967_01Occ-ala-Phe-Gly-Leu-Apr-Leu-Asp-Arg-Ile-NH₂; JAL- 136 1142 B 0974Occ-ala-Phe-Gly-Leu-Eay-Leu-Asp-Arg-Ile-NH₂; JAL- 137 1204 C 0975Occ-ala-Phe-Gly-Leu-Fpr-Leu-Asp-Arg-Ile-NH₂; JAL- 138 1145 C 0978Occ-ala-Phe-Gly-Leu-Dtp-Leu-Asp-Arg-Ile-NH₂; JAL- 139 1174 C 0979Occ-ala-Phe-Gly-Leu-Eaz-Leu-Asp-Arg-Ile-NH₂; JAL- 140 1146 C 0980Occ-Az3-Phe-Gly-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 141 1139 C 0985Occ-ala-Phe-Gly-Leu-Pro-Leu-Asp-Arg-Tbg- JAL- 142 1127 C NH₂; 0989Occ-ala-Phe-Gly-Leu-Pro-Leu-Ser-Arg-Ile-NH₂; JAL- 143 1099 C 0992Occ-ala-Phe-Gly-Leu-Pro-Leu-Hse-Arg-Ile-NH₂; JAL- 144 1113 C 0993Occ-ala-Phe-Gly-Ile-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 145 1127 C 0995Occ-ala-Phe-Gly-Nva-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 146 1113 C 0996Occ-ala-Phe-Gly-Hle-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 147 1141 C 0998Occ-ala-Thi-Gly-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 148 1133 C 1000Occ-ala-Pcf-Gly-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 149 1161 C 1002Occ-ala-Thk-Gly-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 150 1133 C 1003Occ-ala-Mtf-Gly-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 151 1195 C 1005Occ-ala-Mmf-Gly-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 152 1141 C 1006Occ-ala-Phe-ser-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 153 1157 B 1010Occ-ala-Phe-thr-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 154 1171 B 1011Occ-ala-Phe-val-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 155 1169 C 1012Occ-ala-Phe-leu-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 156 1183 B 1013Occ-ala-Phe-nle-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 157 1183 B 1014Occ-Sni-Phe-Gly-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 158 1197 B NH₂; 1015Occ-ala-Phe-Gly-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 159 1157 B NH₂; 1016Occ-ala-Phe-asn-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 160 1184 B 1017Occ-ala-Phe-met-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 161 1201 B 1018Occ-ala-Phe-abu-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 162 1155 B 1019Occ-ala-Phe-dap-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 163 1156 B 1020Occ-Sni-Phe-nle-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 164 1223 B 1021Occ-Sni-Nmf-nle-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 165 1237 B 1022Occ-Sni-Phe-nle-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 166 1239 A 1024Occ-ala-Phe-nle-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 167 1199 B 1025Occ-ala-Phe-leu-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 168 1199 B 1026Occ-ala-Phe-nva-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 169 1185 B 1027Occ-ala-Phe-phe-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 170 1029 B 1028Occ-ala-Phe-ctb-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 171 1244 B 1029Occ-ala-Phe-lys-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 172 1198 B 1030Occ-ala-Phe-arg-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 173 1226 B 1031Occ-ala-Phe-his-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 174 1207 B 1032Ac-Hgl-ala-ala-Phe-Gly-Leu-Pro-Leu-Asp-Arg- JAL- 175 1255 B Ile-NH₂;1033 Ac-hgl-ala-ala-Phe-Gly-Leu-Pro-Leu-Asp-Arg-Ile- JAL- 176 1255 BNH₂; 1034 Occ-pip-Phe-Gly-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 177 1167 C1035 Occ-ala-Phe-Gly-Leu-Pro-Leu-cDR-Ile-NH₂; JAL- 178 1153 C 1037Occ-ala-Phe-Gly-Leu-Bhp-Leu-Asp-Arg-Ile-NH₂; JAL- 179 1234 C 1038Occ-ala-Phe-leu-Leu-Pro-Nml-Asp-Arg-Ile-NH₂; JAL- 180 1196 A 1039Occ-Sni-Phe-leu-Leu-Pro-Nml-Asp-Arg-Ile-NH₂; JAL- 181 1236 A 1040Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 182 1253 A 1041Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 183 1213 A 1042Occ-ala-Pcf-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 184 1247 A 1043Occ-ala-Phe-nle-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 185 1213 A 1044Occ-ala-Phe-Gly-Leu-Pro-Npl-Asp-Arg-Ile-NH₂; JAL- 186 1169 C 1045Occ-ala-Phe-arg-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 187 1242 A 1047Occ-ala-Phe-asp-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 188 1201 C 1048Occ-ala-Phe-glu-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 189 1215 C 1049Occ-ala-Pcf-leu-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 190 1233 A 1050Occ-ala-Pmf-leu-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 191 1213 B 1051Occ-ala-Nmf-leu-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 192 1213 A 1052Occ-pro-Phe-leu-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 193 1225 A 1053Occ-pip-Phe-leu-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 194 1239 A 1054Occ-ala-Phe-lys-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 195 1228 A 1060Occ-ala-Phe-orn-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 196 1214 A 1061Occ-ala-Phe-lys-Leu-Pro-Nml-Asp-Arg-Ile-NH₂; JAL- 197 1212 B 1065Occ-ala-Phe-lys-Leu-Pro-Nml-Ala-Arg-Ile-NH₂; JAL- 198 1168 C 1068Occ-ala-Phe-arg-Leu-Pro-Nml-Asp-Arg-Ile-NH₂; JAL- 199 1240 B 1075Occ-ala-Nmf-arg-Leu-Pro-Nml-Asp-Arg-Ile-NH₂; JAL- 200 1254 B 1076Occ-pip-Nmf-arg-Leu-Pro-Nml-Asp-Arg-Ile-NH₂; JAL- 201 1294 A 1077Occ-pip-Phe-arg-Leu-Pro-Nml-Asp-Arg-Ile-NH₂; JAL- 202 1280 A 1078Occ-ala-Nmf-arg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 203 1270 A NH₂; 1085Occ-ala-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 204 1256 A 1086Occ-pip-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 205 1296 A 1087Occ-ala-Phe-arg-Leu-Tfp-Leu-Asp-Arg-Ile-NH₂; JAL- 206 1244 B 1114Occ-ala-Phe-Gly-Leu-Tfp-Leu-Asp-Arg-Ile-NH₂; JAL- 207 1145 B 1115Occ-ala-Pbf-arg-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 208 1321 A 1116Occ-ala-Phe-dab-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 209 1169 B 1120Occ-ala-Phe-nar-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 210 1212 B 1121Occ-ala-Phe-gdp-Leu-Pro-Leu-Asp-Arg-Ile-NH₂; JAL- 211 1198 B 1122Oct-ala-Phe-arg-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 212 1227 B 1156_02Oct-pip-Phe-arg-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 213 1267 C 1157_02Occ-ala-Phe-arg-(KM-116-167)-Nml-Asp-Arg-Ile- JAL- 214 1226 C NH₂; 1159832-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 215 1241 B 1214Occ-ala-Phe-arg-Leu-Hyp-Ile-Asp-Arg-Ile-NH₂; JAL- 216 1242 B 1224Occ-ala-Phe-arg-Leu-Hyp-Npg-Asp-Arg-Ile-NH₂; JAL- 217 1256 A 1225Occ-ala-Phe-arg-Leu-Hyp-Tbg-Asp-Arg-Ile-NH₂; JAL- 218 1242 C 1226Occ-ala-Phe-arg-Leu-Hyp-Ebe-Asp-Arg-Ile-NH₂; JAL- 219 1246 B 1227Occ-ala-Phe-arg-Leu-Lys-Nml-Asp-Arg-Ile-NH₂; JAL- 220 1271 B 1228Occ-ala-Phe-arg-Leu-Nmk-Nml-Asp-Arg-Ile- JAL- 221 1285 B NH₂; 1229Occ-ala-Phe-arg-Leu-Nma-Nml-Asp-Arg-Ile- JAL- 222 1228 C NH₂; 1230Occ-ala-Phe-arg-Leu-Sar-Nml-Asp-Arg-Ile-NH₂; JAL- 223 1214 B 1231Occ-ala-Phe-arg-Nva-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 224 1242 B 1232Occ-ala-Phe-arg-Ebe-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 225 1260 B 12336014-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 226 1239 B 12376015-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 227 1239 B 12386054-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 228 1241 B 12396056-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 229 1239 B 12406057-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 230 1259 B 12416058-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 231 1259 B 12426059-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 232 1274 B 1243832-Nmf-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 233 1255 C 1244832-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 234 1196 B 1245832-Phe-arg-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 235 1225 C 1246Oct-Sni-FrL-Hyp-Leu-Asp-Arg-Ile-NH₂; JAL- 236 1268 B 1248Occ-ala-Phe-Gly-Leu-Tap-Leu-Asp-Arg-Ile-NH₂; JAL- 237 1142 A 1249Occ-ala-Phe-arg-Leu-Tap-Leu-Asp-Arg-Ile-NH₂; JAL- 238 1241 A 1250Occ-ala-Phe-leu-Leu-Tap-Asp-Arg-Ile-NH₂; JAL- 239 1198 A 1251Occ-ala-Phe-ser-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 240 1187 A 1252Occ-Sni-Phe-ser-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 241 1227 B 1253Occ-Sni-Phe-lys-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 242 1268 A 1254Occ-Sni-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 243 1296 A 1255Occ-Sni-Mpa-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 244 1254 C NH₂; 1256Occ-Sni-Ppa-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 245 1254 C 1257(6071-OH)-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 246 1230 C NH₂; 1259(6072-OH)-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 247 1258 B NH₂; 12605587-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 248 1214 C 1261Occ-ala-Phe-Gly-Leu-Tap(2Me)-Leu-Asp-Arg-Ile- JAL- 249 1170 B NH₂; 1262Occ-ala-Phe-arg-Leu-Tap(2Me)-Leu-Asp-Arg-Ile- JAL- 250 1269 B NH₂; 1263Occ-ala-Phe-leu-Leu-Tap(2Me)-Leu-Asp-Arg-Ile- JAL- 251 1226 B NH₂; 1264Occ-Sni-Phe-orn-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 252 1254 A NH₂; 1265Occ-Sni-Opa-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 253 1254 B NH₂; 1266Occ-ala-Nmf-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 254 1227 A NH₂; 1267Occ-ala-Nmf-lys-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 255 1242 B NH₂; 1268Occ-ala-Nmf-orn-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 256 1228 B NH₂; 1269Occ-ala-Phe-Gly-Leu-Gup-Leu-Asp-Arg-Ile-NH₂; JAL- 257 1184 B 1270Occ-ala-Phe-arg-Leu-Gup-Leu-Asp-Arg-Ile-NH₂; JAL- 258 1283 B 1271Occ-ala-Phe-leu-Leu-Gup-Leu-Asp-Arg-Ile-NH₂; JAL- 259 1240 B 1272Oct-Sar-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 260 1242 B 1273Oct-aze-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 261 1254 B 1274Oct-Az3-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 262 1254 B NH₂; 1275Occ-ala-Phe-leu-Leu-Eal-Nml-Asp-Arg-Ile-NH₂; JAL- 263 1198 B 1281_01Occ-ala-Phe-Gly-Leu-Eal-Nml-Asp-Arg-Ile-NH₂; JAL- 264 1144 C 1282Occ-ala-Phe-leu-Leu-Hyp-(SH-158)-Asp-Arg-Ile- JAL- 265 1227 A NH₂; 1283Occ-ala-Phe-arg-Leu-Hyp-(SH-158)-Asp-Arg-Ile- JAL- 266 1271 A NH₂; 1284Occ-Sni-Phe-dap(Me2)-Leu-Hyp-Nml-Asp-Arg- JAL- 267 1254 A Ile-NH₂; 1287Occ-ala-Phe-orn(Me2)-Leu-Hyp-Nml-Asp-Arg- JAL- 268 1242 A Ile-NH₂; 1288Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Asp-Arg- JAL- 269 1282 A Ile-NH₂; 1289(AR-201-48)-arg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 270 1242 C NH₂; 1291(AR-201-49)-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 271 1257 B NH₂; 1292(AR-201-48)-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 272 1199 C NH₂; 1293(AR-201-49)-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 273 1214 A NH₂; 1294Occ-Sni-Phe-leu-Leu-Tap-Nml-Asp-Arg-Ile-NH₂; JAL- 274 1252 A 1295Occ-ala-Phe-leu-Leu-Tap-Nml-Asp-Arg-Ile-NH₂; JAL- 275 1212 A 1296Oct-Sni-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 276 1282 A 12976182-ala-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 277 1280 B NH₂; 1298Oct-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 278 1239 A 1302Occ-ala-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Tbg- JAL- 279 1256 A NH₂; 1305Occ-ala-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Eca- JAL- 280 1254 B NH₂; 1306Occ-ala-Phe-arg-Leu-Hyp-Dap(Me2)-Asp-Arg-Ile- JAL- 281 1242 B NH₂; 1314Occ-ala-Phe-arg-Dap(Me2)-Hyp-Nml-Asp-Arg- JAL- 282 1257 C Ile-NH₂; 1315(AR-201-54)-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 283 1277 B NH₂; 1316Occ-Sni-Phe-arg-Leu-Tap-Nml-Asp-Arg-Ile-NH₂; JAL- 284 1295 A 1317Occ-Sni-Phe-orn-Leu-Tap-Nml-Asp-Arg-Ile-NH₂; JAL- 285 1253 A 1318Occ-Sni-Phe-nle-Leu-Tap-Nml-Asp-Arg-Ile-NH₂; JAL- 286 1252 B 1319Occ-Sni-Phe-Gly-Leu-Tap-Nml-Asp-Arg-Ile- JAL- 287 1196 A NH₂; 1320Occ-Sni-Phe-leu-Leu-Tap(Ac)-Nml-Asp-Arg-Ile- JAL- 288 1294 B NH₂; 1321Occ-Sni-Phe-leu-Leu-Tap(G)-Nml-Asp-Arg-Ile- JAL- 289 1309 A NH₂; 1322Occ-Sni-Phe-leu-Leu-Tap(Bal)-Nml-Asp-Arg-Ile- JAL- 290 1323 A NH₂; 13236059(O)-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 291 1291 B NH₂; 1324Occ-Sni-Phe-dap(Me2)-Leu-Tap-Nml-Asp-Arg- JAL- 292 1253 A Ile-NH₂; 1325Oct-Sni-Phe-leu-Leu-Tap-Nml-Asp-Arg-Ile-NH₂; JAL- 293 1238 A 1326Occ-ala-Phe-arg-Leu-Hyp-Nml-Asp-Orn-Ile-NH₂; JAL- 294 1214 B 1327Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Orn-Ile-NH₂; JAL- 295 1171 B 1328Occ-ala-Phe-arg-Leu-Hyp-Nml-Glu-Arg-Ile-NH₂; JAL- 296 1270 C 1329Occ-ala-Phe-leu-Leu-Hyp-Nml-Glu-Arg-Ile-NH₂; JAL- 297 1227 B 1330Occ-ala-Phe-arg-Leu-Hyp-Nml-Val-Arg-Ile-NH₂; JAL- 298 1240 B 1331Occ-ala-Phe-leu-Leu-Hyp-Nml-Val-Arg-Ile-NH₂; JAL- 299 1197 A 1332Occ-ala-Phe-leu-Leu-Hyp-Nml-Val-Arg-Ile-NH₂; JAL- 300 1197 B 1332_02Occ-ala-Phe-arg-Leu-Hyp-Nml-Thr-Arg-Ile-NH₂; JAL- 301 1242 B 1333Occ-ala-Phe-leu-Leu-Hyp-Nml-Thr-Arg-Ile-NH₂; JAL- 302 1199 B 1334Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Eca- JAL- 303 1211 B NH₂; 1335Occ-ala-Phe-Fhy-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 304 1240 A NH₂; 1336Occ-ala-Phe-Egg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 305 1254 B NH₂; 1337Occ-ala-Phe-Apc-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 306 1226 A NH₂; 1338(AR-201-58)-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 307 1254 C NH₂; 1339(AR-201-59)-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 308 1267 C NH₂; 1340(AR-201-62)-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 309 1253 B NH₂; 1341(AR-201-69)-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 310 1317 B NH₂; 1342Sbt-Sni-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 311 1309 A 1343Nbt-Sni-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 312 1309 B 1344Sbt-ala-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 313 1269 C 1345Nbt-ala-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 314 1269 C 1346Occ-ala-Phe-dap(Me2)-Leu-Tap-Nml-Asp-Arg- JAL- 315 1213 B Ile-NH₂; 1347Occ-Sni-Phe-leu-Leu-Tap(Et2)-Nml-Asp-Arg-Ile- JAL- 316 1308 B NH₂; 1348Occ-Sni-Phe-leu-Leu-Tap(Et)-Nml-Asp-Arg-Ile- JAL- 317 1280 A NH₂; 1349Occ-ala-Phe-Apc-Leu-Tap-Nml-Asp-Arg-Ile- JAL- 318 1265 A NH₂; 1350Occ-Sni-Phe-Apc-Leu-Tap-Nml-Asp-Arg-Ile- JAL- 319 1265 A NH₂; 1351Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Tbg- JAL- 320 1213 A NH₂; 1352Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Egz- JAL- 321 1225 A NH₂; 1358Occ-ala-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Egz- JAL- 322 1268 B NH₂; 1359Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Nle-Ile-NH₂; JAL- 323 1170 C 1360Occ-ala-Phe-arg-Leu-Hyp-Nml-Asp-Nle-Ile-NH₂; JAL- 324 1213 C 1361Occ-ala-Phe-arg-Leu-Hyp-Nml-Ile-Arg-Ile-NH₂; JAL- 325 1254 C 1362Occ-ala-Phe-leu-Leu-Hyp-Nml-Ile-Arg-Ile-NH₂; JAL- 326 1211 B 1363Occ-ala-Phe-arg-Leu-Hyp-Oic-Asp-Arg-Ile-NH₂; JAL- 327 1280 B 1364Occ-ala-Phe-arg-Leu-Hyp-Pip-Asp-Arg-Ile-NH₂; JAL- 328 1240 C 1365Occ-ala-Phe-leu-Leu-Hyp-Pip-Asp-Arg-Ile-NH₂; JAL- 329 1197 B 1366Occ-ala-Phe-leu-Leu-Hyp-Dap(Me2)-Asp-Arg-Ile- JAL- 330 1200 A NH₂; 1367Occ-ala-Phe-leu-Dap(Me2)-Hyp-Nml-Asp-Arg- JAL- 331 1214 B Ile-NH₂; 1368Oct-Sni-Phe-dap(Me2)-Leu-Tap-Nml-Asp-Arg- JAL- 332 1239 A Ile-NH₂; 1369Occ-Sni-Phe-dap(6263)2-Leu-Tap-Nml-Asp-Arg- JAL- 333 1311 B Ile-NH₂;1370 Occ-Sni-Phe-leu-Leu-Tap(Ae)-Nml-Asp-Arg-Ile- JAL- 334 1295 A NH₂;1371 Occ-Sni-Phe-leu-Leu-Tap(Ap)-Nml-Asp-Arg-Ile- JAL- 335 1309 A NH₂;1372 (AR-201-58)-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 336 1211 B NH₂;1373 (AR-201-62)-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 337 1210 B NH₂;1374 (AR-201-69)-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 338 1274 B NH₂;1375 (AR-201-72)-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 339 1227 C NH₂;1376 (AR-201-72)-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 340 1270 C NH₂;1377 (AR-201-73)-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 341 1216 B NH₂;1378 (AR-201-73)-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 342 1259 B NH₂;1379 (AR-201-68)-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 343 1274 A NH₂;1380 (AR-201-68)-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 344 1317 B NH₂;1381 Sbt-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 345 1266 A 1382Nbt-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 346 1266 B 1383Occ-ala-Phe-leu-Leu-Hyp-Oic-Asp-Arg-Ile-NH₂; JAL- 347 1237 B 1386Occ-ala-Phe-arg-Leu-Hyp-Pro-Asp-Arg-Ile-NH₂; JAL- 348 1226 C 1387Occ-ala-Phe-arg-Leu-Hyp-Aze-Asp-Arg-Ile-NH₂; JAL- 349 1212 C 1393Occ-ala-Phe-arg-Leu-Hyp-Eat-Asp-Arg-Ile-NH₂; JAL- 350 1244 C 1394Occ-ala-Phe-arg-Leu-Hyp-Eaz-Asp-Arg-Ile-NH₂; JAL- 351 1244 C 1395Occ-ala-Phe-arg-Leu-Hyp-Tic-Asp-Arg-Ile-NH₂; JAL- 352 1288 B 1396Occ-Sni-Phe-leu-Leu-Hyp-Nml-Val-Arg-Ile-NH₂; JAL- 353 1237 A 1398Oct-Sni-Phe-leu-Leu-Hyp-Nml-Val-Arg-Ile-NH₂; JAL- 354 1223 B 1399Occ-Sni-Phe-dap(Me2)-Leu-Hyp-Nml-Val-Arg- JAL- 355 1238 C Ile-NH₂; 1400Occ-Sni-Phe-leu-Leu-Tap-Nml-Val-Arg-Ile-NH₂; JAL- 356 1236 A 1401Oct-Sni-Phe-dap(Me2)-Leu-Hyp-Nml-Val-Arg- JAL- 357 1224 B Ile-NH₂; 1402Occ-Sni-Phe-dap(Me2)-Leu-Tap-Nml-Val-Arg- JAL- 358 1237 A Ile-NH₂; 1403Oct-Sni-Phe-leu-Leu-Tap-Nml-Val-Arg-Ile-NH₂; JAL- 359 1222 B 1404Oct-Sni-Phe-dap(Me2)-Leu-Tap-Nml-Val-Arg-Ile- JAL- 360 1223 A NH₂; 1405Occ-ala-Phe-Apc(Me)-Met-glu--Leu-Hyp-Nml- JAL- 361 1240 AAsp-Arg-Ile-NH₂; 1406 Occ-ala-Phe-Apc(Et)-Glu-thr--Leu-Hyp-Nml-Asp- JAL-362 1254 A Arg-Ile-NH₂; 1407 Occ-ala-Phe-Apc(Ae)-Ala-glu--Leu-Hyp-Nml-JAL- 363 1269 B Asp-Arg-Ile-NH₂; 1408Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Aib- JAL- 364 1185 B NH₂; 1413Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Aml- JAL- 365 1227 A NH₂; 1414Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Deg- JAL- 366 1213 A NH₂; 1416Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Nmr-Ile- JAL- 367 1227 A NH₂; 1417Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Pro-Ile-NH₂; JAL- 368 1254 B 1418Occ-ala-Phe-leu-Leu-Hyp-Nml-Tbg-Arg-Ile-NH₂; JAL- 369 1211 C 1420Occ-ala-Phe-leu-Leu-Hyp-Nml-Chg-Arg-Ile-NH₂; JAL- 370 1237 C 1421Occ-ala-Phe-leu-Leu-Hyp-Nml-Cpa-Arg-Ile-NH₂; JAL- 371 1195 C 1424Oct-Sni-Phe-dap(Me2)-Leu-Hyp-Nml-Asp-Arg- JAL- 372 1240 A Ile-NH₂; 1429Miy-Hgl-ala-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 373 1561 A NH₂; 1430Miy-Gab-Hgl-ala-Phe-arg-Leu-Hyp-Nml-Asp- JAL- 374 1647 C Arg-Ile-NH₂;1431 Ac-Miy-Gab-Hgl-ala-Phe-arg-Leu-Hyp-Nml-Asp- JAL- 375 1730 CArg-Ile-NH₂; 1432 Occ-ala-Phe-arg-Leu-Hyp-Nml-Asp-Pro-Ile-NH₂; JAL- 3761198 B 1434 Occ-ala-Phe-Apc-Leu-Hyp-Nml-Asp-Pro-Ile- JAL- 377 1167 BNH₂; 1435 Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Pro-Ile-NH₂; JAL- 378 1194 B1436 Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Aze-Ile-NH₂; JAL- 379 1140 B 1437Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Pip-Ile-NH₂; JAL- 380 1168 B 1438Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Hyp-Ile-NH₂; JAL- 381 1170 C 1441Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Eaz-Ile-NH₂; JAL- 382 1173 B 1442Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Cpp-Ile-NH₂; JAL- 383 1167 B 1443Occ-ala-Phe-leu-Leu-Tap-Nml-Asp-Pro-Ile-NH₂; JAL- 384 1153 B 1450Occ-ala-Phe-Apc-Leu-Hyp-Nml-Asp-Pro-Ile- JAL- 385 1207 B NH₂; 1451Occ-ala-Phe-Apc-Leu-Tap-Nml-Asp-Pro-Ile-NH₂; JAL- 386 1166 A 1452Occ-ala-Phe-dap(Me2)-Leu-Tap-Nml-Asp-Pro-Ile- JAL- 387 1154 A NH₂; 1453Occ-ala-Phe-Egz-Leu-Tap-Nml-Asp-Arg-Ile-NH₂; JAL- 388 1224 A 1454Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Eay-Ile-NH₂; JAL- 389 1230 C 1456Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Egz-Ile-NH₂; JAL- 390 1182 C 1457Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Apc-Ile-NH₂; JAL- 391 1183 B 1458Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Tap-Ile-NH₂; JAL- 392 1169 C 1459Occ-ala-Phe-dap(6238)2-Leu-Tap-Nml-Asp-Arg- JAL- 393 1380 B Ile-NH₂;1460 Occ-ala-Phe-dap(6238)-Leu-Tap-Nml-Asp-Arg- JAL- 394 1282 B Ile-NH₂;1461 Occ-ala-Phe-dap(3846)2-Leu-Tap-Nml-Asp-Arg- JAL- 395 1345 BIle-NH₂; 1462 Occ-ala-Phe-dap(1464)-Leu-Tap-Nml-Asp-Arg- JAL- 396 1255 AIle-NH₂; 1463 Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Pro-558; JAL- 397 1162 B1464 Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Pro-Ile-OH; JAL- 398 1194 C 1474Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Pro-Ile-(NH—CH₃); JAL- 399 1207 B 1475Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Chy-Ile-NH₂; JAL- 400 1170 B 1476Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-H3p-Ile-NH₂; JAL- 401 1170 B 1477Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Dhp-Ile-NH₂; JAL- 402 1152 B 1479Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Udp-Ile-NH₂; JAL- 403 1143 B 1482Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Bhk-Ile-NH₂; JAL- 404 1199 B 1483Occ-Sni-Nif-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 405 1298 B 1486Occ-Sni-Pff-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 406 1271 A 1487Occ-Sni-Pmy-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 407 1283 B NH₂; 1488Occ-Sni-Tyr-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 408 1269 C 1489Occ-Sni-Bmf-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 409 1267 C NH₂; 1490Occ-Sni-Eay-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 410 1279 B 1491Occ-Sni-Paf-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 411 1268 B 1492Occ-Sni-Pcf-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 412 1287 A 1493Occ-Sni-Pmf-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 413 1267 A NH₂; 1494Occ-Sni-Eaa-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 414 1322 A 1496Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Pro-2118; JAL- 415 1210 B 1506Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Pro-2906; JAL- 416 1134 C 1508Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Pro-1381; JAL- 417 1164 B 1509Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Pro-1381; JAL- 418 1164 B 1509_02Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Pro-1860; JAL- 419 1176 A 1510Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Pro-1906; JAL- 420 1174 B 1511Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Pro-Che; JAL- 421 1176 A 1512_02Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Pro-5121; JAL- 422 1178 C 1513Occ-Sni-Phe-Ala-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 423 1211 C NH₂; 1553Occ-Sni-Phe-Leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 424 1253 B NH₂; 1554Occ-Sni-Phe-Apc-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 425 1266 A NH₂; 1555Occ-ala-Phe-leu-Leu-Hyp-Nml-(BB725)-Arg-Ile- JAL- 426 1224 A NH₂; 1556Occ-ala-Phe-leu-Leu-Hyp-Nml-(BB726)-Arg-Ile- JAL- 427 1238 C NH₂; 1557Occ-ala-Phe-leu-Leu-Hyp-Nml-(BB727)-Arg-Ile- JAL- 428 1238 A NH₂; 1558Occ-Sni-Phe-leu-Leu-Tap-Nml-Asp-Pro-Ile-NH₂; JAL- 429 1194 A 1559Occ-Sni-Phe-Gly-Leu-Tap-Nml-Asp-Pro-Ile-NH₂; JAL- 430 1138 B 1560Occ-Sni-Phe-Apc-Leu-Tap-Nml-Asp-Pro-Ile- JAL- 431 1207 A NH₂; 1561Occ-Sni-Phe-leu-Leu-Tap-Nml-Asp-Pro-Che; JAL- 432 1176 A 1568Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Nmi- JAL- 433 1267 A NH₂; 1569Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Nmr-Ile- JAL- 434 1267 B NH₂; 1570Occ-Sni-Phe-leu-Nml-Hyp-Nml-Asp-Arg-Ile- JAL- 435 1267 C NH₂; 1572Occ-Sni-Nmf-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 436 1267 A NH₂; 1573Occ-Sni-Phe-nml-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 437 1267 C NH₂; 1574Occ-Sni-Phe-dap(Me2)-Leu-Hyp-Nml-Asp-Arg- JAL- 438 1268 C Nmi-NH₂; 1575Occ-Sni-Phe-dap(Me2)-Leu-Hyp-Nml-Asp-Nmr- JAL- 439 1268 A Ile-NH₂; 1576Occ-Sni-Phe-dap(Me2)-Leu-Hyp-Nml-Nmd-Arg- JAL- 440 1268 C Ile-NH₂; 1577Occ-Sni-Phe-dap(Me2)-Nml-Hyp-Nml-Asp-Arg- JAL- 441 1268 B Ile-NH₂; 1578Occ-Sni-Nmf-dap(Me2)-Leu-Hyp-Nml-Asp-Arg- JAL- 442 1268 A Ile-NH₂; 1579Occ-Sni-Phe-dap(Me2)-Leu-Hyp-Nml-Asp-Pro- JAL- 443 1195 A Ile-NH₂; 1580Occ-Sni-Phe-leu-Leu-Hyp-Nml-Val-Pro-Che; JAL- 444 1160 B 1594Occ-Sni-Phe-leu-Leu-Hyp-Npg-Asp-Pro-Che; JAL- 445 1177 A 1595Occ-Sni-Phe-leu-Leu-Hyp-Ile-Asp-Pro-Che; JAL- 446 1162 B 1596Occ-Sni-Nmf-leu-Leu-Hyp-Nml-Asp-Pro-Che; JAL- 447 1191 A 1597Occ-Sni-Phe-leu-Nml-Hyp-Nml-Asp-Pro-Che; JAL- 448 1190 C 1598Occ-Sni-Eaa-leu-Leu-Hyp-Nml-Asp-Pro-Che; JAL- 449 1245 A 1599Occ-Sni-Phe-Gly-Leu-Hyp-Nml-Asp-Pro-Che; JAL- 450 1120 B 1600Occ-Sni-Phe-Apc-Leu-Hyp-Nml-Asp-Pro-Che; JAL- 451 1190 B 1601Occ-Sni-Phe-Apc-Leu-Tap-Nml-Asp-Pro-Che; JAL- 452 1190 A 1602Occ-Sni-Phe-dap(Me2)-Leu-Tap-Nml-Asp-Pro- JAL- 453 1177 A Che; 1603Occ-Sni-Phe-dap(Me2)-Leu-Hyp-Nml-Asp-Pro- JAL- 454 1177 A Che; 16041319-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 455 1272 A NH₂; 16051320-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 456 1286 A NH₂; 16062553-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 457 1302 C NH₂; 16074734-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 458 1316 B NH₂; 16094703-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 459 1339 B NH₂; 16126988-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 460 1342 C NH₂; 1615Hex-(3421)-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg- JAL- 461 1360 B Ile-NH₂;1616 1695-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 462 1372 C NH₂; 1617Occ-Sni-Mcf-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 463 1287 A NH₂; 1618Occ-Sni-Pbf-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 464 1332 A 1619Occ-Sni-Thk-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 465 1259 A NH₂; 1620Occ-Sni-Mtf-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 466 1321 A 1621Occ-Sni-Otf-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 467 1321 C 1622Occ-Sni-Phe-ctb-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 468 1299 A 1623Occ-Sni-Phe-leu-Nle-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 469 1253 A 1624Occ-Sni-Phe-leu-Leu-Hyp-Ile-Asp-Arg-Ile-NH₂; JAL- 470 1239 A 1625Occ-Sni-Phe-leu-Leu-Hyp-Cpg-Asp-Arg-Ile-NH₂; JAL- 471 1251 A 1626Occ-Sni-Phe-leu-Leu-Hyp-Chg-Asp-Arg-Ile-NH₂; JAL- 472 1265 B 1627Occ-Sni-NPhe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 473 1253 C NH₂; 1634Occ-Sni-NHfe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 474 1267 C NH₂; 1635Occ-(aFL)-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 475 1225 B 1636Occ-(afL)-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 476 1225 B 1637Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Eaz-Che; JAL- 477 1195 A 1638Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Eal-Che; JAL- 478 1177 B 1639Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-(ES-283- JAL- 479 1163 B 049); 1646Occ-Sni-Phe-leu-Leu-Hyp-Nml-Glu-Pro-Che; JAL- 480 1191 B 1652Occ-Sni-Phe-leu-Leu-Tap-Nml-Val-Pro-Che; JAL- 481 1160 A 1654Occ-Sni-Phe-leu-Nle-Hyp-Nml-Asp-Pro-Che; JAL- 482 1177 A 1657779-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 483 1263 B 1659785-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 484 1335 C 16601281-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 485 1259 B NH₂; 16613218-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 486 1293 C NH₂; 16646013-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 487 1285 B NH₂; 16655587-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 488 1281 A NH₂; 16661281-G-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 489 1316 C NH₂; 16681281-Bal-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 490 1330 C NH₂; 1669Occ-(AFL)-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 491 1225 A 1671Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Apc-Che; JAL- 492 1204 C 1672Occ-Sni-Phe-leu-Leu-Hyp-Nml-NP-Che; JAL- 493 1176 C 1673Occ-Sni-Phe-leu-Leu-Tap-Nml-(BB726)-Pro-Che; JAL- 494 1200 B 1676Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Pca-Che; JAL- 495 1192 A 1679Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Che; JAL- 496 1236 A 1680Occ-Sni-Phe-leu-Leu-Tap(Ae)-Nml-Asp-Arg- JAL- 497 1278 A Che; 1681Occ-Sni-Phe-leu-Leu-Tap-Nml-Asp-Arg-Che; JAL- 498 1235 A 1682Occ-Sni-Phe-leu-Leu-Tap(Ac)-Nml-Val-Arg-Che; JAL- 499 1262 C 1683Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Apc(Gua)- JAL- 500 1248 C Che; 1685Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Apc(Gly)- JAL- 501 1263 B Che; 1687Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-(BB394)- JAL- 502 1166 C Che; 1694Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-(BB785)- JAL- 503 1192 B Che; 1697Occ-Sni-Hfe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 504 1267 C NH₂; 1701Occ-ala-Nmf-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 505 1227 C NH₂; 1702Occ-Sni-Phe-leu-Leu-Tap-Nml-Val-Arg-Che; JAL- 506 1218 A 1729Occ-Sni-Phe-leu-Leu-Hyp-Nml-Val-Arg-Che; JAL- 507 1219 A 1730Occ-Sni-Phe-leu-Leu-Hyp-Nml-Ala-Arg-Che; JAL- 508 1193 C 1750Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asn-Arg-Che; JAL- 509 1236 C 1751Occ-Sni-Phe-leu-Leu-Hyp-Nml-Ser-Arg-Che; JAL- 510 1209 A 1752Occ-Sni-Phe-leu-Leu-Hyp-Nml-Thr-Arg-Che; JAL- 511 1223 A 1753Occ-Sni-Phe-leu-Leu-Hyp-Nml-Nle-Arg-Che; JAL- 512 1235 C 1755Occ-Sni-Phe-leu-Leu-Hyp-Nml-Ble-Arg-Che; JAL- 513 1235 B 1756Occ-Sni-Phe-leu-Leu-Hyp-Nml-Thi-Arg-Che; JAL- 514 1275 C 1758Occ-Sni-Phe-leu-Leu-Hyp-Nml-Chg-Arg-Che; JAL- 515 1261 C 1763(AR-314-87)-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 516 1279 A NH₂; 1765-2(AR-314-102)-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 517 1239 A NH₂; 1774Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Asp-Arg- JAL- 518 1265 A Che; 1776Occ-Sni-Phe-lys(Me2)-Leu-Hyp-Nml-Asp-Arg- JAL- 519 1279 A Che; 1777Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Val-Arg- JAL- 520 1249 B Che; 1778Occ-Sni-Phe-lys(Me2)-Leu-Hyp-Nml-Val-Arg- JAL- 521 1263 A Che; 1779Occ-Sni-Phe-lys(Me2)-Leu-Hyp-Nml-Asp-Arg- JAL- 522 1296 B Ile-NH₂; 1781Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Val-Arg- JAL- 523 1266 A Ile-NH₂; 1782Occ-Sni-Phe-lys(Me2)-Leu-Hyp-Nml-Val-Arg-Ile- JAL- 524 1280 C NH₂; 1783Occ-Sni-Phe-dab(Me2)-Leu-Hyp-Nml-Val-Arg- JAL- 525 1235 A Che; 1784Occ-Sni-Phe-dab(Me2)-Leu-Hyp-Nml-Asp-Arg- JAL- 526 1268 A Ile-NH₂; 1785Occ-Sni-Phe-dab(Me2)-Leu-Hyp-Nml-Val-Arg- JAL- 527 1252 B Ile-NH₂; 1786Occ-Nhpr-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 528 1257 B NH₂; 1798Occ-Nbhp-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 529 1273 B NH₂; 1799Occ-ser-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 530 1229 B 1800Occ-hse-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 531 1243 B 1801Gluc-Aoa-Hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp- JAL- 532 1503 B Arg-Ile-NH₂;1802 Gluc-Aoa-hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp- JAL- 533 1503 AArg-Ile-NH₂; 1803 (1913)-Hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg- JAL- 5341384 B Ile-NH₂; 1804 (1270)-Hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg- JAL-535 1396 C Ile-NH₂; 1805 (1888)-Hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-JAL- 536 1428 B Ile-NH₂; 1806Occ-Hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 537 1394 C NH₂; 1807H-Adx-Hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg- JAL- 538 1413 A Ile-NH₂; 18081888-hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 539 1428 B NH₂; 1837H-Adx-hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg- JAL- 540 1413 B Ile-NH₂; 1838Oct-Sni-Phe-leu-Leu-Tap-Nml-Asp-Arg-Che; JAL- 541 1221 A 1843Oct-Sni-Phe-leu-Leu-Tap-Nml-Val-Pro-Che; JAL- 542 1146 B 1844Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Val-Pro- JAL- 543 1190 B Che; 1845Occ-Sni-Phe-orn(Me2)-Leu-Tap-Nml-Val-Pro- JAL- 544 1189 B Che; 1846Oct-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Val-Pro- JAL- 545 1176 C Che; 1847Oct-Sni-Phe-leu-Leu-Hyp-Nml-Val-Arg-Che; JAL- 546 1206 B 1848Oct-Sni-Phe-leu-Leu-Tap-Nml-Val-Arg-Che; JAL- 547 1205 B 1849Occ-Sni-Phe-orn(Me2)-Leu-Tap-Nml-Val-Arg- JAL- 548 1248 A Che; 1850Oct-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Val-Arg- JAL- 549 1235 B Che; 1851Occ-Sni-Phe-leu-Leu-Hyp-Nml-Bmf-Arg-Ile- JAL- 550 1299 C NH₂; 1857Occ-Sni-Phe-leu-Leu-Hyp-Nml-Phg-Arg-Ile-NH₂; JAL- 551 1859 C 1858Occ-Sni-Phe-leu-Leu-Hyp-Nml-Cpg-Arg-Ile-NH₂; JAL- 552 1263 B 1859Occ-Sni-Phe-leu-Leu-Hyp-Nml-(AR-314-145)- JAL- 553 1277 C Arg-Ile-NH₂;1864 (AR-314-169)-Phe-leu-Leu-Hyp-Nml-Asp-Arg- JAL- 554 1281 B Ile-NH₂;1868-2 (AR-314-170)-Phe-leu-Leu-Hyp-Nml-Asp-Arg- JAL- 555 1253 CIle-NH₂; 1869-2 (AR-314-171)-Phe-leu-Leu-Hyp-Nml-Asp-Arg- JAL- 556 1281C Ile-NH₂; 1870-2 (AR-385-008)-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 5571273 C NH₂; 1873 (AR-314-172)-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 558 1287B NH₂; 1874 Occ-Sni-Phe-(AR-385-12)-Leu-Hyp-Nml-Asp- JAL- 559 1294 AArg-Ile-NH₂; 1877 Occ-Sni-Phe-hse-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 560 1241B NH₂; 1878 Occ-Sni-Phe-abu(pip)-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 561 1308B NH₂; 1879 (AR-385-042)-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 562 1287 BNH₂; 1880 Occ-Sni-Phe-Fbz-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 563 1280 B NH₂;1881 Occ-Sni-Phe-Fhy-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 564 1280 B NH₂; 1882Occ-Sni-Phe-thr-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 565 1241 C 1883Occ-Sni-Phe-his-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; JAL- 566 1277 B 1884Occ-Sni-Phe-metO2-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 567 1303 B NH₂; 1885Occ-Sni-Phe-(AR-385-017)-Leu-Hyp-Nml-Asp- JAL- 568 1310 B Arg-Ile-NH₂;1886 Occ-Sni-Phe-opa-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 569 1288 B NH₂; 1887Occ-Sni-Phe-mpa-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 570 1288 B NH₂; 1888Occ-Sni-Phe-ppa-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 571 1288 B NH₂; 1889Occ-Sni-Phe-Egg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 572 1294 A NH₂; 1890Occ-Sni-Phe-Eao-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 573 1299 B NH₂; 1892Occ-Sni-Phe-Aic-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 574 1299 B NH₂; 1893Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Ser-Arg- JAL- 575 1237 B Che; 1894Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Thr-Arg- JAL- 576 1251 A Che; 1895H-Hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 577 1268 B NH₂; 1896H-hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 578 1268 B NH₂; 1897H-Lys-Hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg- JAL- 579 1396 B Ile-NH₂; 1898H-Lys-hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg- JAL- 580 1396 B Ile-NH₂; 1899H-Lys-Pro-Hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp- JAL- 581 1493 A Arg-Ile-NH₂;1900 (2857-Ac)-Hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp- JAL- 582 1489 BArg-Ile-NH₂; 1901 (1625-Ac)-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg- JAL- 5831268 B Ile-NH₂; 1907 Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Dim-Arg- JAL- 5841264 B Ile-NH₂; 1910 Occ-Sni-Phe-leu-Leu-Hyp-Nml-Pse-Arg-Ile-NH₂; JAL-585 1305 C 1912 Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Pth-Arg-Ile- JAL- 5861348 C NH₂; 1913 Occ-Sni-Phe-Dha-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 587 1209B NH₂; 1915_2 Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Pse-Arg- JAL- 588 1316 CChe; 1916 Occ-Sni-Phe-leu-Leu-Hyp-Nml-Pse-Arg-Che; JAL- 589 1288 C 1917Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Pth-Arg- JAL- 590 1330 B Che; 1918Occ-Sni-Phe-leu-Leu-Hyp-Nml-Pth-Arg-Che; JAL- 591 1302 B 1919Occ-Sni-Phe-leu-Leu-Hyp-Nml-Ser-Arg-Ile-NH₂; JAL- 592 1225 B 1920Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Ser-Arg-Ile- JAL- 593 1254 C NH₂; 1921Occ-Sni-Phe-leu-Leu-Hyp-Nml-Cya-Arg-Ile-NH₂; JAL- 594 1289 B 1922Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Cya-Arg- JAL- 595 1318 B Ile-NH₂; 1923Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Thr-Arg- JAL- 596 1268 B Ile-NH₂; 1924Occ-Sni-Phe-leu-Leu-Hyp(Asp(—))-Nml-Asp-Arg- JAL- 597 1368 B Ile-NH₂;1928 Occ-Sni-Phe-leu-Leu-Hyp(2581)-Nml-Asp-Arg- JAL- 598 1338 B Ile-NH₂;1929 Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-OH; JAL- 599 1254 B 1930Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Asp-Arg- JAL- 600 1283 B Ile-OH; 1931Occ-Sni-Phe-leu-Leu-Hyp-Nml-Thr-Arg-Ile-NH₂; JAL- 601 1239 A 1932Occ-Sni-Phe-leu-Leu-Tap(Asp(—))-Nml-Asp-Arg- JAL- 602 1367 B Ile-NH₂;1935 Occ-Sni-Phe-orn(Me2)-Leu-Tap-Nml-Asp-Arg- JAL- 603 1281 A Ile-NH₂1936

Preferred NPR-B agonists of the present invention are those peptideswithin activity group B, as presented in Table 3, above. Most preferredNPR-B agonists of the present invention are those peptides withinactivity group A, as presented in Table 4, below.

TABLE 4 Most preferred compounds according to the present invention andtheir agonistic activity in in vitro assays. SEQ ID (M + H)⁺ in ActivityStructure JAL NO: MS [amu] (group)Occ-Sni-Phe-nle-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂ JAL- 166 1239 A 1024Occ-ala-Phe-leu-Leu-Pro-Nml-Asp-Arg-Ile-NH₂ JAL- 180 1196 A 1039Occ-Sni-Phe-leu-Leu-Pro-Nml-Asp-Arg-Ile-NH₂ JAL- 181 1236 A 1040Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 182 1253 A 1041Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 183 1213 A 1042Occ-ala-Pcf-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 184 1247 A 1043Occ-ala-Phe-nle-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 185 1213 A 1044Occ-ala-Phe-arg-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂ JAL- 187 1242 A 1047Occ-ala-Pcf-leu-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂ JAL- 190 1233 A 1050Occ-ala-Nmf-leu-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂ JAL- 192 1213 A 1052Occ-pro-Phe-leu-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂ JAL- 193 1225 A 1053Occ-pip-Phe-leu-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂ JAL- 194 1239 A 1054Occ-ala-Phe-lys-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 195 1228 A 1060Occ-ala-Phe-orn-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 196 1214 A 1061Occ-pip-Nmf-arg-Leu-Pro-Nml-Asp-Arg-Ile-NH₂ JAL- 201 1294 A 1077Occ-pip-Phe-arg-Leu-Pro-Nml-Asp-Arg-Ile-NH₂ JAL- 202 1280 A 1078Occ-ala-Nmf-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 203 1270 A 1085Occ-ala-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 204 1256 A 1086Occ-pip-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 205 1296 A 1087Occ-ala-Pbf-arg-Leu-Hyp-Leu-Asp-Arg-Ile-NH₂ JAL- 208 1321 A 1116Occ-ala-Phe-arg-Leu-Hyp-Npg-Asp-Arg-Ile-NH₂ JAL- 217 1256 A 1225Occ-ala-Phe-Gly-Leu-Tap-Leu-Asp-Arg-Ile-NH₂ JAL- 237 1142 A 1249Occ-ala-Phe-arg-Leu-Tap-Leu-Asp-Arg-Ile-NH₂ JAL- 238 1241 A 1250Occ-ala-Phe-leu-Leu-Tap-Asp-Arg-Ile-NH₂ JAL- 239 1198 A 1251Occ-ala-Phe-ser-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 240 1187 A 1252Occ-Sni-Phe-lys-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 242 1268 A 1254Occ-Sni-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 243 1296 A 1255Occ-Sni-Phe-orn-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 252 1254 A 1265Occ-ala-Nmf-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 254 1227 A 1267Occ-ala-Phe-leu-Leu-Hyp-(SH-158)-Asp-Arg-Ile- JAL- 265 1227 A NH₂ 1283Occ-ala-Phe-arg-Leu-Hyp-(SH-158)-Asp-Arg-Ile- JAL- 266 1271 A NH₂ 1284Occ-Sni-Phe-dap(Me2)-Leu-Hyp-Nml-Asp-Arg- JAL- 267 1254 A Ile-NH₂ 1287Occ-ala-Phe-orn(Me2)-Leu-Hyp-Nml-Asp-Arg- JAL- 268 1242 A Ile-NH₂ 1288Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Asp-Arg- JAL- 269 1282 A Ile-NH₂ 1289(AR-201-49)-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 273 1214 A NH₂ 1294Occ-Sni-Phe-leu-Leu-Tap-Nml-Asp-Arg-Ile-NH₂ JAL- 274 1252 A 1295Occ-ala-Phe-leu-Leu-Tap-Nml-Asp-Arg-Ile-NH₂ JAL- 275 1212 A 1296Oct-Sni-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 276 1282 A 1297Oct-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 278 1239 A 1302Occ-ala-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Tbg-NH₂ JAL- 279 1256 A 1305Occ-Sni-Phe-arg-Leu-Tap-Nml-Asp-Arg-Ile-NH₂ JAL- 284 1295 A 1317Occ-Sni-Phe-orn-Leu-Tap-Nml-Asp-Arg-Ile-NH₂ JAL- 285 1253 A 1318Occ-Sni-Phe-Gly-Leu-Tap-Nml-Asp-Arg-Ile-NH₂ JAL- 287 1196 A 1320Occ-Sni-Phe-leu-Leu-Tap(G)-Nml-Asp-Arg-Ile- JAL- 289 1309 A NH₂ 1322Occ-Sni-Phe-leu-Leu-Tap(Bal)-Nml-Asp-Arg-Ile- JAL- 290 1323 A NH₂ 1323Occ-Sni-Phe-dap(Me2)-Leu-Tap-Nml-Asp-Arg- JAL- 292 1253 A Ile-NH₂ 1325Oct-Sni-Phe-leu-Leu-Tap-Nml-Asp-Arg-Ile-NH₂ JAL- 293 1238 A 1326Occ-ala-Phe-leu-Leu-Hyp-Nml-Val-Arg-Ile-NH₂ JAL- 299 1197 A 1332Occ-ala-Phe-Fhy-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 304 1240 A 1336Occ-ala-Phe-Apc-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 306 1226 A 1338Occ-Sni-Phe-leu-Leu-Tap(Et)-Nml-Asp-Arg-Ile- JAL- 317 1280 A NH₂ 1349Occ-ala-Phe-Apc-Leu-Tap-Nml-Asp-Arg-Ile-NH₂ JAL- 318 1265 A 1350Occ-Sni-Phe-Apc-Leu-Tap-Nml-Asp-Arg-Ile-NH₂ JAL- 319 1265 A 1351Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Tbg-NH₂ JAL- 320 1213 A 1352Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Egz-NH₂ JAL- 321 1225 A 1358Occ-ala-Phe-leu-Leu-Hyp-Dap(Me2)-Asp-Arg-Ile- JAL- 330 1200 A NH₂ 1367Oct-Sni-Phe-dap(Me2)-Leu-Tap-Nml-Asp-Arg- JAL- 332 1239 A Ile-NH₂ 1369Occ-Sni-Phe-leu-Leu-Tap(Ae)-Nml-Asp-Arg-Ile- JAL- 334 1295 A NH₂ 1371Occ-Sni-Phe-leu-Leu-Tap(Ap)-Nml-Asp-Arg-Ile- JAL- 335 1309 A NH₂ 1372(AR-201-68)-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 343 1274 A NH₂ 1380Sbt-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 345 1266 A 1382Occ-Sni-Phe-leu-Leu-Hyp-Nml-Val-Arg-Ile-NH₂ JAL- 353 1237 A 1398Occ-Sni-Phe-leu-Leu-Tap-Nml-Val-Arg-Ile-NH₂ JAL- 356 1236 A 1401Occ-Sni-Phe-dap(Me2)-Leu-Tap-Nml-Val-Arg- JAL- 358 1237 A Ile-NH₂ 1403Oct-Sni-Phe-dap(Me2)-Leu-Tap-Nml-Val-Arg-Ile- JAL- 360 1223 A NH₂ 1405Occ-ala-Phe-Apc(Me)-Met-glu--Leu-Hyp-Nml- JAL- 361 1240 AAsp-Arg-Ile-NH₂ 1406 Occ-ala-Phe-Apc(Et)-Glu-thr--Leu-Hyp-Nml-Asp- JAL-362 1254 A Arg-Ile-NH₂ 1407 Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Aml-NH₂JAL- 365 1227 A 1414 Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Deg-NH₂ JAL-366 1213 A 1416 Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Nmr-Ile-NH₂ JAL- 3671227 A 1417 Oct-Sni-Phe-dap(Me2)-Leu-Hyp-Nml-Asp-Arg- JAL- 372 1240 AIle-NH₂ 1429 Miy-Hgl-ala-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 373 1561A NH₂ 1430 Occ-ala-Phe-Apc-Leu-Tap-Nml-Asp-Pro-Ile-NH₂ JAL- 386 1166 A1452 Occ-ala-Phe-dap(Me2)-Leu-Tap-Nml-Asp-Pro-Ile- JAL- 387 1154 A NH₂1453 Occ-ala-Phe-Egz-Leu-Tap-Nml-Asp-Arg-Ile-NH₂ JAL- 388 1224 A 1454Occ-ala-Phe-dap(1464)-Leu-Tap-Nml-Asp-Arg- JAL- 396 1255 A Ile-NH₂ 1463Occ-Sni-Pff-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 406 1271 A 1487Occ-Sni-Pcf-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 412 1287 A 1493Occ-Sni-Pmf-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 413 1267 A 1494Occ-Sni-Eaa-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 414 1322 A 1496Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Pro-1860 JAL- 419 1176 A 1510Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Pro-Che JAL- 421 1176 A 1512_02Occ-Sni-Phe-Apc-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 425 1266 A 1555Occ-ala-Phe-leu-Leu-Hyp-Nml-(BB725)-Arg-Ile- JAL- 426 1224 A NH₂ 1556Occ-ala-Phe-leu-Leu-Hyp-Nml-(BB727)-Arg-Ile- JAL- 428 1238 A NH₂ 1558Occ-Sni-Phe-leu-Leu-Tap-Nml-Asp-Pro-Ile-NH₂ JAL- 429 1194 A 1559Occ-Sni-Phe-Apc-Leu-Tap-Nml-Asp-Pro-Ile-NH₂ JAL- 431 1207 A 1561Occ-Sni-Phe-leu-Leu-Tap-Nml-Asp-Pro-Che JAL- 432 1176 A 1568Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Nmi- JAL- 433 1267 A NH₂ 1569Occ-Sni-Nmf-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 436 1267 A 1573Occ-Sni-Phe-dap(Me2)-Leu-Hyp-Nml-Asp-Nmr- JAL- 439 1268 A Ile-NH₂ 1576Occ-Sni-Nmf-dap(Me2)-Leu-Hyp-Nml-Asp-Arg- JAL- 442 1268 A Ile-NH₂ 1579Occ-Sni-Phe-dap(Me2)-Leu-Hyp-Nml-Asp-Pro- JAL- 443 1195 A Ile-NH₂ 1580Occ-Sni-Phe-leu-Leu-Hyp-Npg-Asp-Pro-Che JAL- 445 1177 A 1595Occ-Sni-Nmf-leu-Leu-Hyp-Nml-Asp-Pro-Che JAL- 447 1191 A 1597Occ-Sni-Eaa-leu-Leu-Hyp-Nml-Asp-Pro-Che JAL- 449 1245 A 1599Occ-Sni-Phe-Apc-Leu-Tap-Nml-Asp-Pro-Che JAL- 452 1190 A 1602Occ-Sni-Phe-dap(Me2)-Leu-Tap-Nml-Asp-Pro- JAL- 453 1177 A Che 1603Occ-Sni-Phe-dap(Me2)-Leu-Hyp-Nml-Asp-Pro- JAL- 454 1177 A Che 16041319-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 455 1272 A 16051320-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 456 1286 A 1606Occ-Sni-Mcf-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 463 1287 A 1618Occ-Sni-Pbf-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 464 1332 A 1619Occ-Sni-Thk-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 465 1259 A 1620Occ-Sni-Mtf-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 466 1321 A 1621Occ-Sni-Phe-ctb-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 468 1299 A 1623Occ-Sni-Phe-leu-Nle-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 469 1253 A 1624Occ-Sni-Phe-leu-Leu-Hyp-Ile-Asp-Arg-Ile-NH₂ JAL- 470 1239 A 1625Occ-Sni-Phe-leu-Leu-Hyp-Cpg-Asp-Arg-Ile-NH₂ JAL- 471 1251 A 1626Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Eaz-Che JAL- 477 1195 A 1638Occ-Sni-Phe-leu-Leu-Tap-Nml-Val-Pro-Che JAL- 481 1160 A 1654Occ-Sni-Phe-leu-Nle-Hyp-Nml-Asp-Pro-Che JAL- 482 1177 A 16575587-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 488 1281 A 1666Occ-(AFL)-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 491 1225 A 1671Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Pca-Che JAL- 495 1192 A 1679Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Che JAL- 496 1236 A 1680Occ-Sni-Phe-leu-Leu-Tap(Ae)-Nml-Asp-Arg-Che JAL- 497 1278 A 1681Occ-Sni-Phe-leu-Leu-Tap-Nml-Asp-Arg-Che JAL- 498 1235 A 1682Occ-Sni-Phe-leu-Leu-Tap-Nml-Val-Arg-Che JAL- 506 1218 A 1729Occ-Sni-Phe-leu-Leu-Hyp-Nml-Val-Arg-Che JAL- 507 1219 A 1730Occ-Sni-Phe-leu-Leu-Hyp-Nml-Ser-Arg-Che JAL- 510 1209 A 1752Occ-Sni-Phe-leu-Leu-Hyp-Nml-Thr-Arg-Che JAL- 511 1223 A 1753(AR-314-87)-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 516 1279 A 1765-2(AR-314-102)-leu-Leu-Hyp-Nml-Asp-Arg-Ile- JAL- 517 1239 A NH₂ 1774Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Asp-Arg- JAL- 518 1265 A Che 1776Occ-Sni-Phe-lys(Me2)-Leu-Hyp-Nml-Asp-Arg- JAL- 519 1279 A Che 1777Occ-Sni-Phe-lys(Me2)-Leu-Hyp-Nml-Val-Arg- JAL- 521 1263 A Che 1779Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Val-Arg- JAL- 523 1266 A Ile-NH₂ 1782Occ-Sni-Phe-dab(Me2)-Leu-Hyp-Nml-Val-Arg- JAL- 525 1235 A Che 1784Occ-Sni-Phe-dab(Me2)-Leu-Hyp-Nml-Asp-Arg- JAL- 526 1268 A Ile-NH₂ 1785H-Adx-Hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg- JAL- 538 1413 A Ile-NH₂ 1808Oct-Sni-Phe-leu-Leu-Tap-Nml-Asp-Arg-Che JAL- 541 1221 A 1843Occ-Sni-Phe-orn(Me2)-Leu-Tap-Nml-Val-Arg- JAL- 548 1248 A Che 1850Occ-Sni-Phe-(AR-385-12)-Leu-Hyp-Nml-Asp- JAL- 559 1294 A Arg-Ile-NH₂1877 Occ-Sni-Phe-Egg-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂ JAL- 572 1294 A 1890Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Thr-Arg- JAL- 576 1251 A Che 1895H-Lys-Pro-Hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp- JAL- 581 1493 A Arg-Ile-NH₂1900 Occ-Sni-Phe-leu-Leu-Hyp-Nml-Thr-Arg-Ile-NH₂ JAL- 601 1239 A 1932Occ-Sni-Phe-orn(Me2)-Leu-Tap-Nml-Asp-Arg- JAL- 603 1281 A Ile-NH₂ 1936B. Diseases to be Treated and/or Prevented

The present invention is also directed to methods of treating orpreventing diseases in a subject that involve administering to thesubject a therapeutically effective amount of a composition thatincludes one or more NPR-B agonists as described herein, wherein thedisease is one of the following. The subject may be a mammal, such as ahuman, a primate, a cow, a horse, a dog, a cat, a mouse, or a rat. Inparticular embodiments, the subject is a human.

1. Definitions

“Treatment” and “treating” refer to administration or application of adrug to a subject or performance of a procedure or modality on a subjectfor the purpose of obtaining a therapeutic benefit of a disease orhealth-related condition. The term “therapeutic benefit” used throughoutthis application refers to anything that promotes or enhances thewell-being of the subject with respect to the medical treatment of hiscondition. This includes, but is not limited to, a reduction in thefrequency or severity of the signs or symptoms of a disease. Therapeuticbenefit also includes reducing the signs or symptoms associated withglaucoma in a subject with glaucoma. For example, a therapeutic benefitin a patient with glaucoma is obtained where there is no furtherprogression of visual field loss in the affected eye, or a slowing ofthe rate of progression of visual field loss in the affected eye, or animprovement in vision.

A “disease” or “health-related condition” can be any pathologicalcondition of a body part, an organ, or a system resulting from anycause, such as infection, trauma, genetic defect, age-relateddeterioration of bodily functions, and/or environmental stress. Thecause may or may not be known. Examples of diseases include glaucoma,retinopathies, ocular trauma, and optic neuropathies. Thus, one of skillin the art realizes that a treatment may improve the disease condition,but may not be a complete cure for the disease.

The terms “prevention” and “preventing” are used herein according totheir ordinary and plain meaning to mean “acting before” or such an act.In the context of a particular disease or health-related condition,those terms refer to administration or application of an agent, drug, orremedy to a subject or performance of a procedure or modality on asubject for the purpose of blocking or minimizing the onset of a diseaseor health-related condition. For example, an individual with an eye thatis at risk of developing glaucoma (such as an individual with ocularhypertension) can be treated with a NPR-B agonist as set forth hereinfor the purpose of blocking or minimizing the onset of the signs orsymptoms of glaucoma (i.e., prevention of glaucoma). In a specificembodiment, prevention pertains to lowering elevated intraocularpressure, blocking detectable optic nerve damage as a result of glaucomain a subject, reducing the rate of vision loss in a subject, or haltingloss of vision in a subject. The subject can be a subject who is knownor suspected of being free of a particular disease or health-relatedcondition at the time the relevant preventive agent is administered. Thesubject, for example, can be a subject with no known disease orhealth-related condition (i.e., a healthy subject). In some embodiments,the subject had a previous disease that has been treated in the past andis now known or suspected to be disease-free.

For those skilled in the art it is easy to understand, that differentdiseases are summarized under certain terms or generic terms. Thesesummaries are no limitation and each disease can be viewed on its ownand can be treated or prevented with the compounds according to thepresent invention.

2. Glaucoma and Ocular Hypertension

Glaucoma is the second leading cause of blindness world-wide (Thyleforsand Negrel 1994, Bull World Health Organ. 72:323-326). Open-angleglaucoma (OAG) and angle closure glaucoma combined represent the secondleading cause of blindness worldwide (Quigley and Broman, 2006 Br JOphthalmol. 90:262-267). Angle-closure glaucoma is more common in theAsian population (Foster et al. 2000, Arch Ophthalmol. 118:1105-11),while open-angle glaucoma is more commonly found in black patients(Leske et al. 2007, Ophthalmic Epidemiol. 14:166-172). Glaucoma is aprogressive disease in which the risk of vision loss increases withdisease duration. In light of an aging population world-wide, the impactof this blinding disorder can be expected to increase in the future.

The disease state referred to as glaucoma is a family of diseasescharacterized by a permanent loss of visual function due to irreversibledamage to the optic nerve. More specifically, glaucoma results in opticneuropathy leading to the loss of retinal ganglion cell (RGC) functionfollowed by apoptotic cell death and a progressive increase in visionloss. Morphologically or functionally distinct types of glaucoma aretypically characterized by elevated intraocular pressure (IOP), which isconsidered to be an important risk factor of the pathological course ofthe disease. Disruption of normal aqueous outflow leading to elevatedIOP is integral to glaucoma pathophysiology. Ocular hypertension is acondition wherein IOP is elevated but no apparent loss of visualfunction has occurred; such patients are considered to be at high riskfor the eventual development of the visual loss associated withglaucoma. Some patients with glaucomatous field loss have relatively lowIOPs. These so called normotension or low tension glaucoma patients canalso benefit from agents that lower and control IOP.

Glaucoma is typically identified by changes in IOP, visual fielddeficits and/or fundus changes at the optic disk. Elevated IOP, found inmost glaucoma patients, is a result of morphological and biochemicalchanges in the trabecular meshwork (TM), an aqueous humor filteringtissue located at the iris-cornea angle of the eye. As glaucomaprogresses, there is a loss of TM cells and a buildup of extracellularproducts which inhibit the normal aqueous humor outflow resulting in IOPelevation. In addition to elevated IOP, other factors, such as geneticdefects, may lead to mechanical distortion of the optic nerve head (ONH)ultimately resulting in ONH cupping and loss of RGC and their axons. Theexact mechanism of this pathological process is currently unknown. Ithas been suggested that lowering the TOP of patients diagnosed withglaucoma by at least 20-30% will decrease the progressive worsening ofthe disease by 50-60% (Quigley 2005 Ophthalmology 112:1642-1643).Without proper diagnosis and treatment, glaucoma can progress to totalirreversible blindness.

Initially, most open-angle glaucoma patients are managed with one ormore of a wide variety of topical ocular or oral hypotensive medicationsthat act to increase aqueous fluid outflow and/or decrease aqueous fluidproduction, or with surgical procedures such as laser trabeculoplastyand filtration surgery. Treatment regimens currently available forpatients exhibiting elevated IOP, regardless of cause, typically includethe topical application, from once daily to multiple times per day, ofone or multiple eyedrops or pills containing a small moleculeTOP-lowering compound. Also, pills that decrease the amount of aqueoushumor created can be given between two and four times daily. Glaucomamedications typically prescribed include cholinergic agonists,adrenergic agonists, beta adrenergic blockers, carbonic anhydraseinhibitors and prostaglandin analogs. Although these classes ofmedications are effective in controlling IOP, each of them has certainlimitations in efficacy and untoward effects. For example, betaadrenergic blockers do not lower IOP at night; many glaucoma patients donot respond to a particular drug class; and a majority of glaucomapatients require the use of a combination of drugs. In addition, many ofthe drugs cause local irritation of the eye, such as burning, stinging,itching, tearing, conjunctival hyperemia, foreign body sensation,blurred vision, and eye pain. Some occasionally induce systemic sideeffects. Hence, there is a genuine and continuous need for novel andimproved glaucoma medications.

“Glaucoma” and “glaucomatous optic neuropathy” and “glaucomatousretinopathy,” as used herein, are interchangeable. Glaucoma refers to adisease characterized by the permanent loss of visual function due toirreversible damage to the retinal ganglion cells in the retina andoptic nerve. The major risk factor for glaucoma and the related loss ofvisual function is elevated intraocular pressure. There are differenttypes of glaucoma, including primary open angle glaucoma (POAG), angleclosure glaucoma, and congenital/developmental glaucoma.

As used herein, the term “intraocular pressure” or “IOP” refers to thepressure of the content inside the eye. In a normal human eye, IOP istypically in the range of 10 to 21 mm Hg. IOP varies among individuals,for example, it may become elevated due to anatomical problems,inflammation of the eye, as a side-effect from medication or due togenetic factors. “Elevated” intraocular pressure is currently consideredto be ≧21 mm Hg, which is also considered to be a major risk factor forthe development of glaucoma.

However, some individuals with an elevated IOP may not develop glaucomaand are considered to have ocular hypertension. “Ocular hypertension” asused herein refers to a condition in which the intraocular pressure inthe eye of a subject is higher than normal but the optic nerve andvisual fields are within normal limits. These individuals may besusceptible to developing the loss of visual function that is typicallyassociated with glaucoma. As used herein, the terms “susceptible,” or“susceptibility” refers to an individual or subject that is or at riskof developing optic nerve damage or retinal damage that is associatedwith elevated intraocular pressure.

Thus, the present invention is directed to methods of treating orpreventing an ophthalmic disease in a subject that involve administeringto the subject a therapeutically effective amount of a composition thatincludes one or more NPR-B agonists as described herein, wherein theophthalmic disease is glaucoma, elevated intraocular pressure or ocularhypertension. The subject may be a mammal, such as a human, a primate, acow, a horse, a dog, a cat, a mouse, or a rat. In particularembodiments, the subject is a human.

In preferred aspects, the NPR-B agonists of the invention will lowerintraocular pressure associated with glaucoma. The glaucoma may be anytype of glaucoma, such as primary open angle glaucoma, angle closureglaucoma, normal tension glaucoma, congenital glaucoma, neovascularglaucoma, steroid-induced glaucoma, or glaucoma related to ocular trauma(e.g., ghost cell glaucoma or glaucoma related to choroidal detachment).

The present invention is also directed to methods of loweringintraocular pressure in a subject, comprising administering to thesubject a pharmaceutically effective amount of a composition comprisinga NPR-B agonist described herein, wherein intraocular pressed islowered. In particular embodiments, the subject is a human. For example,in specific embodiments, the human is a patient with ocular hypertensionor elevated IOP.

3. CNP Deficiencies as in Diabetes

Diabetic nephropathy is a progressive kidney disease, resulting fromlongstanding diabetes mellitus. Experimental evidence shows thatnatriuretic peptides play a pathophysiological role in the glomerularabnormalities seen in diabetes mellitus. BNP overexpression preventeddiabetic nephropathy in a streptozotocin-induced mouse model of diabetes(Makino et al. 2006, Diabetologia. 49:2514-2524). In another study withstreptozotocin-induced diabetic rats, cardiac CNP mRNA concentrationswere decreased 2.6-fold (Walther et al. 2000, J Mol Endocrinol.24:391-395). In a genetic model of diabetes, the non-obese diabeticmouse, mesangial cells derived from diabetic mice showed constitutiveoverexpression of NPR-C; this was associated with a reduced response ofcGMP production to ANP or CNP treatment (Ardaillou et al. 1999, KidneyInt 55:1293-1302).

4. Conditions with Hyperproliferation of Vascular Smooth Muscle Cells

The abnormal growth of vascular smooth muscle cells (VSMC) is a commoncause of many vascular diseases. A disturbance of the balance betweengrowth inhibitors and growth promoters results in the hyperproliferationof those cells, and vasoactive substances, including natriureticpeptides, seem to play a major role in this process. Early experimentalfindings indicate that the guanylyl-cyclase-linked natriuretic peptidereceptors mediate anti-proliferative activity of the natriureticpeptides on vascular smooth muscle cell growth (Hutchinson et al. 1997,Cardiovasc Res. 35:158-167). Ex vivo experiments showed a directinhibition of growth in rat VSMCs by CNP (Furuya et al. 1991, BiochemBiophys Res Commun. 177:927-931). Furthermore, migration of rat VSMCscould be inhibited by CNP (Ikeda et al. 1997, Arterioscler Thromb VascBiol. 17:731-736). CNP gene transfer resulted in a reduction of the VSMCproliferation in pig femoral arteries in vivo, and the effect was evensuperior over CNP peptide application (Pelisek et al. 2006, J Gene Med.8:835-844). In another report, CNP gene transfer resulted in thesuppression of vascular remodeling in porcine coronary arteries in vivo(Morishige et al. 2000, J Am Coll Cardiol. 35:1040-1047), thus furtherstrengthening the rationale of using CNP to offset thehyperproliferation of VSMCs.

5. Cardiac Pathologies, Especially Heart Failure and Hypertrophy

Considerable evidence supports a central pathophysiological role fornatriuretic peptides in cardiovascular diseases, and in particular heartfailure. The advantage of focusing on CNP in this indication is theunchanged reactivity of NPR-B, while NPR-A activity was shown to bereduced in this condition (Dickey et al. 2007, Endocrinology.148:3518-3522, Nakamura et al. 1994, Circulation. 90:1210-1214). Thefact that plasma CNP is elevated in heart failure patients (Del Ry etal. 2005, Eur J Heart Fail. 7:1145-1148, Del Ry et al. 2007, Peptides.28:1068-1073) is interpreted as part of a compensatory vasodilatingresponse in the peripheral vasculature (Del Ry et al. 2005, Eur J HeartFail. 7:1145-1148, Wright et al. 2004, Hypertension. 43:94-100).Traditional treatment of heart failure aims at the support of cardiacfunction by preventing cardiomyocyte loss and hypertrophy. CNP is ableto support cardiac function via a positive effect on the vitality ofcardiomyocytes (Rosenkranz et al. 2003, Cardiovasc Res. 57:515-522,Tokudome et al. 2004, Endocrinology. 145:2131-2140). Also, CNP reducedcardiac fibrosis (Horio et al. 2003, Endocrinology. 144:2279-2284), theeffect being stronger than that by ANP or BNP. Results from studies ondogs showed a potential inotropic effect of CNP (Beaulieu et al. 1997,Am J Physiol. 273:H1933-1940), supporting the potential of CNP to treatheart failure.

Hypertrophy of the heart is an enlargement of the organ, due to anincrease in the volume of its muscular fibres. Experimental evidencesuggests that CNP exhibits important autocrine and paracrine functionswithin the heart and the coronary circulation (D'Souza et al. 2004,Pharmacol Ther. 101:113-129). In vivo administration of CNP has beenshown to improve cardiac function and attenuate cardiac remodeling aftermyocardial infarction in rats (Soeki et al. 2005, J Am Coll Cardiol45:608-616). Another recent study shows that CNP is able to reducereactive hypertrophy of cardiomyocytes after an experimental myocardialinfarction in transgenic mice over-expressing CNP in cardiomyocytes(Wang et al. 2007, Eur J Heart Fail. 9:548-557).

6. Cardiovascular Pathologies, Especially Atherosclerosis, Hypertension,Endothelial Dysfunction and Thrombotic Events

Atherosclerosis is a chronic inflammatory response in the walls ofarterial blood vessels. In vitro evidence suggests that CNP has aninhibitory role in vascular smooth muscle cell proliferation andmigration (Furuya et al. 1991, Biochem Biophys Res Commun. 177:927-931,Shinomiya et al. 1994, Biochem Biophys Res Commun. 205:1051-1056).Type-C natriuretic peptide inhibited neointimal thickening in injuredarteries of rabbits and rats in vivo (Furuya et al. 1995, Ann N Y AcadSci. 748:517-523, Ueno et al. 1997, Circulation. 96:2272-2279). In anexperimental model of atherosclerosis in rabbits, local infusion of CNPresulted in the preservation of endothelial function and the preventionof neointimal thickening, which normally results from endothelial injury(Gaspari et al. 2000, Clin Exp Pharmacol Physiol. 27:653-655).

Pulmonary hypertension is a progressive disease, characterized by anelevated pressure in the pulmonary arterial system. Common treatment isthe use of vasodilatory substances. The ability of CNP to relaxarteries, possibly via direct interaction with the VSMCs, has been showbefore in isolated pig coronary arteries (Marton et al. 2005, VasculPharmacol. 43:207-212). More specifically, CNP was able to amelioratemonocrotaline-induced pulmonary hypertension in rats and improvedsurvival (Itoh et al. 2004, Am J Respir Crit Care Med. 170:1204-1211),even if treatment with CNP started 3 weeks after the onset of symptoms.

Endothelial dysfunction plays a fundamental role in the development ofatherosclerosis and restenosis. In a rabbit model with features similarto those of the early stage of atherosclerosis or restenosis, chronicperi-arterial administration of ANP or CNP prevented endothelialdysfunction and development of neointima (Gaspari et al. 2000, Clin ExpPharmacol Physiol. 27:653-655, Barber et al. 2005, J Vasc Res.42:101-110).

Prevention of thrombotic events is critical to the management ofcardiovascular diseases. The anti-thrombotic effect of CNP is well known(Ahluwalia et al. 2004, Basic Res Cardiol. 99:83-89). Thrombus formationwas significantly suppressed in the presence of CNP in antilogous rabbitjugular vein grafts (Ohno et al. 2002, Circulation. 105:1623-1626). In amodel of balloon-injured rabbit carotid arteries CNP was shown to exertanti-thrombotic activity, probably via an increase in the NO productionby enhancing the expression of inducible NO synthase (Qian et al. 2002,Circ Res 91:1063-1069).

7. Stimulation of Arteriogenesis

Arteriogenesis refers to the growth of collateral arterioles intofunctional collateral arteries, and is linked to elevated bloodpressure, and elevated flow, causing shear stress against the wall ofthe arterioles. The stimulation of this event presents a strategy totreat arterial occlusive diseases (van Royen et al. 2001, CardiovascRes. 49:543-553). A beneficial effect of ANP on coronary collateralblood flow has been shown earlier (Kyriakides et al. 1998, Clin Cardiol.21:737-742).

8. Inflammation, Especially Reduction of Inflammatory Mediators, e.g.TNF-Alpha, Other Cytokines or any Kind of Inflammatory Mediator

Several publications suggest a role of CNP in the modulation ofinflammatory responses: in a model of balloon-injured rabbit carotidarteries, in vivo expression of CNP lowered the expression of theinflammatory marker ICAM-1, and reduced the infiltration of macrophages,supposedly via enhancement of NO generation (Qian et al. 2002, Circ Res91:1063-1069). In another study, in rat aortic smooth muscle cells invitro, CNP augmented the transcriptional activation of iNOS induced byinflammatory cytokines (interleukin-1 and tumour necrosis factor-α) andhence the production of NO (Marumo et al. 1995, Endocrinology.136:2135-2142). CNP infusion in rats with an acute experimentalmyocarditis led to a reduction of CD68-positive inflammatory cellinfiltration, and lowered myocardial and serum levels of monocytechemoattractant protein-1 (Obata et al. 2007, Biochem Biophys ResCommun. 356:60-66). By selectively attenuating the expression ofP-selectin, CNP suppressed leukocyte rolling induced by IL-1β orhistamine in a rapid, reversible, and concentration-dependent manner inmice (Scotland et al. 2005, Proc Natl Acad Sci USA. 102:14452-14457). Ina model of bleomycin-induced pulmonary fibrosis in mice, infusion of CNPmarkedly reduced bronchoalveolar lavage fluid IL-1β levels (Murakami etal. 2004, Am J Physiol Lung Cell Mol Physiol. 287:L1172-1177).

9. Pathological Leukocyte Adhesion to Endothelium and Diapedesis IntoTissue

In mouse mesenteric postcapillary venules in vivo in animals with highbasal leukocyte activation (endothelial nitric oxide synthase knockoutmice) or under acute inflammatory conditions (induced by IL-1β orhistamine), CNP suppressed basal leukocyte rolling in a rapid,reversible, and concentration-dependent manner. CNP was also able toinhibit platelet-leukocyte interactions (Scotland et al. 2005, Proc NatlAcad Sci USA. 102:14452-14457). In a model of bleomycin-inducedpulmonary fibrosis in mice, infusion of CNP for 14 days significantlyinhibited infiltration of macrophages into the alveolar and interstitialregions (Murakami et al. 2004, Am J Physiol Lung Cell Mol Physiol.287:L1172-1177). CNP is also known to lower the expression of celladhesion molecules such as ICAM-1 (Qian et al. 2002, Circ Res91:1063-1069), and P-Selectin (Scotland et al. 2005, Proc Natl Acad SciUSA. 102:14452-14457), further strengthening its role in adhesionmolecule modulation.

10. Kidney Disease, Especially Renal Insufficiency, Renal Failure Due ToReduced Renal Perfusion, Glomerulonephritis and Kidney Fibrosis

Local CNP production and CNP receptor expression have previously beendemonstrated in glomeruli (Terada et al. 1994, Am J Physiol.267:F215-222, Lohe et al. 1995, J Am Soc Nephrol. 6:1552-1558, Mattinglyet al. 1994, Kidney Int. 46:744-747, Dean et al. 1994, Am J Physiol.266:F491-496), in kidney cells (Zhao et al. 1994, Kidney Int.46:717-725) and in mesangial cells (Suga et al. 1992, Hypertension.19:762-765), suggesting a role in kidney physiology. In severalconditions CNP levels in plasma or urine are altered. CNP in plasma andurine was increased in nephrotic syndrome (Cataliotti et al. 2002, Am JPhysiol Renal Physiol 283:F464-472), CNP was increased in urine incirrhosis with renal impairment (Gulberg et al. 2000, Gut. 47:852-857),renal and urine levels of CNP were increased in experimental diabetes(Shin et al. 1998, J Endocrinol. 158:35-42), and NP levels were elevatedin chronic kidney disease, but decreased after hemodialysis ortransplantation (Horl 2005, J Investig Med 53:366-370).

The benefit from using CNP in indications such as renal insufficiency,and renal failure, comes from its ability to relax smooth muscles inconduit arteries (Drewett et al. 1995, J Biol Chem. 270:4668-4674,Madhani et al. 2003, Br J Pharmacol. 139:1289-1296), venodilation (Chenand Burnett 1998, J Cardiovasc Pharmacol. 32 Suppl 3:S22-28, Wei et al.1993, J Clin Invest. 92:2048-2052), and dilation of both, afferent andefferent arterioles in glomeruli, as shown in the hydronephrotic ratkidney (Endlich and Steinhausen 1997, Kidney Int. 52:202-207).

Glomerulopathies like glomerulonephritis are typically associated withmesangial cell proliferation, and leukocyte infiltration (Buschhausen etal. 2001, Cardiovasc Res. 51:463-469). The inhibitory effect of CNP onleukocyte infiltration via downregulation of ICAM-1 has been shownbefore (Qian et al. 2002, Circ Res 91:1063-1069, Buschhausen et al.2001, Cardiovasc Res. 51:463-469). In addition, all NPs showanti-proliferative effects on mesangial cells in vitro on rat cells(Suganami et al. 2001, J Am Soc Nephrol 12:2652-2663). In vivo, CNPinfusion improved immune mediated glomerulonephritis in a ratmesangioproliferative anti-Thy 1.1 model (Canaan-Kuhl et al. 1998,Kidney Int 53:1143-1151). In yet another study CNP inhibited glomerularmesangial cell proliferation, MCP-1 secretion, and reduced collagen IVproduction from mesangial cells (Osawa et al. 2000, Nephron.86:467-472).

The inhibitory effect of CNP on the proliferation of glomerularmesangial cells (Suganami et al. 2001, J Am Soc Nephrol 12:2652-2663,Canaan-Kuhl et al. 1998, Kidney Int 53:1143-1151, Osawa et al. 2000,Nephron. 86:467-472) suggests its use in the treatment of kidneyfibrosis.

11. Liver Diseases, Especially Portal Vein Hypertension, LiverCirrhosis, Liver Ascites, Liver Fibrosis and Hepatorenal Syndrome

Evidence for a local natriuretic peptide system in the human liver comesfrom mRNA analysis; specific transcripts for all three NPRs, namelyNPR-A, NPR-B, and NPR-C, could be detected, along with mRNA for ANP andCNP, but not BNP (Vollmar et al. 1997, Gut. 40:145-150). During chronicliver diseases, hepatic stellate cells, believed to play a role in thepathogenesis of liver fibrosis and portal hypertension (Friedman 1993, NEngl J Med. 328:1828-1835), acquire a myofibroblastic phenotype,proliferate, and synthetize components associated with fibrosis.Activation of NPR-B by CNP in myofibroblastic hepatic stellate cells wasshown to inhibit both growth and contraction (Tao et al. 1999, J BiolChem. 274:23761-23769), suggesting that during chronic liver diseases,CNP may counteract both liver fibrogenesis and associated portalhypertension.

Liver cirrhosis is the result of a chronic liver disease characterizedby replacement of liver tissue by fibrous scar tissue. The presence ofCNP in the human kidney and urine (Mattingly et al. 1994, Kidney Int.46:744-747) suggests a role for CNP in fluid and electrolytehomeostasis, and thus possibly a role in renal function disturbances inpatients with cirrhosis of the liver. CNP in the urine of cirrhoticpatients with impaired renal function was increased, while plasma levelswere normal (Gulberg et al. 2000, Gut. 47:852-857). In cirrhoticpatients, ANP infusion reduced the portal pressure and increased thehepatic blood flow, indicative of a lowering of intra-hepatic resistanceto portal flow (Brenard et al. 1992, J Hepatol. 14:347-356).Administration of pharmacological doses of CNP to cirrhotic ratssignificantly decreased portal pressure and peripheral vascularresistance, and increased cardiac output (Komeichi et al. 1995, JHepatol. 22:319-325).

Many disorders can cause ascites, but cirrhosis is the most common.Hence, treatment of disorders such as liver cirrhosis will eventuallyhelp in the avoidance of ascites.

According to the vasodilation theory, the hepatorenal syndrome is theresult of the effect of vasoconstrictor systems acting on the renalcirculation. Due to this increased activity of the vasoconstrictorsystems, renal perfusion and glomerular filtration rate are markedlyreduced, while tubular function is preserved. Any substance thatincreases renal perfusion and/or glomerular filtration rate is thussuited to be used against the hepatorenal syndrome.

12. Lung Diseases, Especially Pulmonary Hypertension, Asthma andPulmonary Fibrosis

CNP was shown to be locally synthesized in pulmonary tissues andtherefore might have action on airway patency (Suga et al. 1992, CircRes. 71:34-39). In vitro CNP was one order of magnitude more potent thanANP in cGMP production in cultured aortic smooth muscle cells.

Pulmonary hypertension is a progressive disease, characterized by anelevated pressure in the pulmonary arterial system. Common treatment isthe use of vasodilatory substances. The ability to relax arteries,probably via direct interaction with the VSMCs, has been shown before inisolated pig coronary arteries (Marton et al. 2005, Vascul Pharmacol.43:207-212). More specifically, CNP was able to amelioratemonocrotaline-induced pulmonary hypertension in rats and to improvesurvival (Itoh et al. 2004, Am J Respir Crit Care Med. 170:1204-1211),even if treatment with CNP started 3 weeks after the onset of symptoms.

In an ovalbumin-induced asthmatic guinea pig model CNP was able tosignificantly inhibit the bronchoconstriction and microvascular leakagein a dose-dependent manner (Ohbayashi et al. 1998, Eur J Pharmacol.346:55-64). In vivo in asthmatics Fluge et al. could demonstratedose-dependent bronchodilating properties of intravenous natriureticpeptide (Fluge et al. 1995, Regul Pept. 59:357-370).

In a model of bleomycin-induced pulmonary fibrosis in mice, infusion ofCNP markedly attenuated the fibrosis, as indicated by significantdecreases in Ashcroft score and lung hydroxyproline content (Murakami etal. 2004, Am J Physiol Lung Cell Mol Physiol. 287:L1172-1177).Immunohistochemistry on lung sections revealed a significantly reducedinfiltration of macrophages into the alveolar and interstitial regions.The markedly decreased number of Ki-67-positive cells in fibroticlesions of the lung further supports the notion of CNP'santi-proliferative effects on pulmonary fibrosis.

13. Male and Female Fertility Problems, Especially Erectile Dysfunction,Stimulation of Male Fertility and Stimulation of Female Fertility

Penile erection depends on relaxation of the smooth muscle of the corpuscavernosum, one of the sponge-like regions of erectile tissue. Thepresence of NPR-B in rat and rabbit cavernosal membrane was shown by Kimet al. (Kim et al. 1998, J Urol. 159:1741-1746). They also showed thatCNP could trigger the production of cGMP in this tissue, and that CNPwas much more potent than BNP and ANP in doing so. NPR-B was also shownto be located in the human corpus cavernosum penis; in organ bathstudies with corpus cavernosum muscle strips CNP at concentrations of0.1 nM to 1 μM led to smooth muscle relaxation from 5% to 40% (Kuthe etal. 2003, J Urol. 169:1918-1922); further support for a role of CNP inerectile dysfunction comes from a recent study, showing that CNP levelsare associated with the presence, severity, and duration of erectiledysfunction (Vlachopoulos et al. 2008, Eur Urol. in press).

The rationale for using CNP to stimulate male fertility is based on itspotential function in testicular blood supply, the modulation of germcell development and spermatozoan motility, and its role in penileerection (as described above). CNP has been found in seminal plasma ofseveral species (Hosang and Scheit 1994, DNA Cell Biol. 13:409-417,Chrisman et al. 1993, J Biol Chem. 268:3698-3703); human Leydig cells,located adjacent to the seminiferous tubules in the testicle, containboth, CNP and the NPR-B receptor (Middendorff et al. 1996, J ClinEndocrinol Metab. 81:4324-4328). CNP was able to increase testosteronelevels in vitro in purified mouse Leydig cells (Khurana and Pandey 1993,Endocrinology. 133:2141-2149), as well as in vivo in the spermatic veinin men (Foresta et al. 1991, J Clin Endocrinol Metab. 72:392-395).Because testosterone activates the initiation, processing andmaintenance of spermatogenesis, CNP has thus an immediate influence onspermatogenesis. Local injection of natriuretic peptides in vivo in ratscaused a dose-related increase in testicular blood flow (Collin et al.1997, Int J Androl. 20:55-60).

A function of CNP in fertilization, pregnancy and embryonic developmentwas first proposed after the detection of CNP in porcine seminal plasma(Chrisman et al. 1993, J Biol Chem. 268:3698-3703). Further studiesshowed expression of NPR-A and —B receptors in human placenta (Itoh etal. 1994, Biochem Biophys Res Commun. 203:602-607), and their modulationin rat ovary and uterus by the estrous cycle (Huang et al. 1996, Am JPhysiol. 271:H1565-1575, Dos Reis et al. 1995, Endocrinology.136:4247-4253, Noubani et al. 2000, Endocrinology. 141:551-559). Inmice, uterine CNP mRNA concentrations increased during pregnancy,whereas in the ovaries these levels decreased compared to non-pregnantcontrols (Stepan et al. 2001, Regul Pept. 102:9-13). In human placentaand myometrium CNP is expressed with no dependency on gestational age inthe third trimester. Pregnancies with intra-uterine growth retardationshowed an opposite regulation of CNP in placenta and myometrium,indicating an organ-specific function of the peptide in humanreproductive tissue (Stepan et al. 2002, Fetal Diagn Ther. 17:37-41).This could be substantiated by studying NPR-B knock-out mice; femalemice were infertile due to the failure of the female reproductive tractto develop (Tamura et al. 2004, Proc Natl Acad Sci USA.101:17300-17305).

14. Pre-Eclampsia and/or Preterm Labor

Pre-eclampsia, a hypertensive disorder of pregnancy, is usuallyassociated with raised blood pressure, and affects about 2-8% ofpregnancies. Inadequate blood supply to the placenta leads toendothelial dysfunction, eventually resulting in damage to the maternalendothelium and kidney and liver. In severe pre-eclampsia BNP levels areelevated, which might reflect ventricular stress and/or subclinicalcardiac dysfunction associated with the condition (Resnik et al. 2005,Am J Obstet Gynecol. 193:450-454). Pregnancies with intra-uterine growthretardation or pre-eclampsia showed an opposite regulation of CNP, witha decrease in the placenta and an increase in the myometrium comparedwith normal pregnancies (Stepan et al. 2002, Fetal Diagn Ther.17:37-41), while maternal CNP plasma levels remained constant; thiscould indicate a compensatory or causative organ-specific function ofthe peptide in human reproductive tissue under these pathophysiologicalconditions, suggesting that application of CNP may have benefits.

15. Skeletal Growth Disturbances, Especially Decreased Body Height(Dwarfism)

Dwarfism can be caused by over 200 separate medical conditions. C-typenatriuretic peptide, acting through its receptor, NPR-B, plays acritical role in longitudinal bone growth (Olney 2006, Growth Norm IGFRes. 16 Suppl A:S6-14), as it stimulates endochondrial ossification(Tamura et al. 2004, Proc Natl Acad Sci USA. 101:17300-17305, Miyazawaet al. 2002, Endocrinology. 143:3604-3610). A spontaneous autosomalrecessive point mutation in the CNP gene, called long bone abnormality(lbab), causes severe dwarfism in mice (Yoder et al. 2008, Peptides.29:1575-1581, Tsuji et al. 2008, Biochem Biophys Res Commun.376:186-190). Complete absence of CNP in mice resulted in dwarfism andearly death (Chusho et al. 2001, Proc Natl Acad Sci USA. 98:4016-4021).

16. Defects of FGF-R (Fibroblast Derived Growth Factor Receptor)Signaling, Especially Overactivity of FGF-R, or Deficiency of CNP orOsteocrin, or Reduced Level of CNP or Osteocrin in the Growth Plates ofLong Bones

In vitro and ex vivo studies showed that CNP acts within the growthplate. CNP, most likely synthetised by proliferating chondrocytes(Chusho et al. 2001, Proc Natl Acad Sci USA. 98:4016-4021), acts locallyto stimulate further proliferation. As opposing element, the FGF/FGFR-3pathway is known to negatively regulate endochondral ossification viaactivation of the Erk MAP kinase pathway, thus inhibiting chondrocyteproliferation and cartilage matrix production (Krejci et al. 2005, JCell Sci. 118:5089-5100). The targeted overexpression of CNP inchondrocytes offset dwarfism in a mouse model of achondroplasia withactivated fibroblast growth factor receptor 3 in the cartilage,suggesting a direct interaction of their signaling pathways (Yasoda etal. 2004, Nat Med. 10:80-86). Moreover, Ozasa et al. found that CNP wasable to antagonize the activation of the MAPK cascade by FGFs, makingthe CNP/NPR-B pathway attractive as a novel therapeutic target in thetreatment of achondroplasia (Ozasa et al. 2005, Bone. 36:1056-1064). CNPalso partially antagonized the FGF2-induced expression, release andactivation of several matrix-remodeling molecules including severalmatrix metalloproteinases. Independent of FGF signaling, CNP stimulatedthe upregulation of matrix production (Krejci et al. 2005, J Cell Sci.118:5089-5100).

Osteocrin is a specific ligand of the natriuretic peptide clearancereceptor NPR-C that modulates bone growth (Thomas et al. 2003, J BiolChem. 278:50563-50571). By blocking the clearance function of NPR-C, itcauses the local elevation of CNP levels, resulting in the proliferationof chondrocytes (Moffatt et al. 2007, J Biol Chem. 282:36454-36462).

In summary, there is a strong rationale to use CNP in order tocompensate for overactive FGF receptors, and for deficiencies or reducedlevels of CNP or osteocrin.

17. Arthritis, especially Degenerative Diseases of Cartilage Tissue,Osteoarthritis and Cartilage Degeneration and Arthritis in Response toTraumatic Cartilage Injury

The rationale for the use of natriuretic peptides for the treatmentand/or prevention of arthritic diseases comes from the observation thatCNP is involved in the skeletal growth, especially in the generation ofcartilage extracellular matrix (Chusho et al. 2001, Proc Natl Acad SciUSA. 98:4016-4021, Yasoda et al. 2004, Nat Med. 10:80-86), which is ableto stabilize damaged cartilage.

CNP depletion was shown to result in impaired bone growth, like thatobserved in achondroplastic bones, with a similar histological pictureof decreased width in both the proliferative and hypertrophicchondrocyte layers of the growth plate (Chusho et al. 2001, Proc NatlAcad Sci USA. 98:4016-4021). The targeted overexpression of CNP inchondrocytes counteracted dwarfism in a mouse model of achondroplasiawith activated fibroblast growth factor receptor 3 in the cartilage. CNPcorrected the decreased extracellular matrix synthesis in the growthplate through inhibition of the MAPK pathway of FGF signaling, resultingin the stimulation of glucosaminoglycans and cartilage collagen (typeII) synthesis (Yasoda et al. 2004, Nat Med. 10:80-86).

In rat chondrosarcoma chondrocytes, after FGF2-mediated growth arrest,CNP mediated the inhibition of MMP induction, and stimulatedextracellular matrix synthesis (Krejci et al. 2005, J Cell Sci.118:5089-5100, Ozasa et al. 2005, Bone. 36:1056-1064), both effectsresulting in a net increase in cartilage extracellular matrix (Krejci etal. 2005, J Cell Sci. 118:5089-5100).

18. Tissue Engineering and Cartilage Regeneration, Especially for the ExVivo Expansion of Cartilage Cells to a Cell Number Sufficient toTransplant Cells Back into a Patient

CNP has stimulatory activity on glucosaminoglycan and cartilage collagen(type II) synthesis in chondrocytes (Krejci et al. 2005, J Cell Sci.118:5089-5100, Yasoda et al. 2004, Nat Med. 10:80-86), a feature that isbeneficial for in vivo regeneration of cartilage. To produce ex vivotissue from the limited number of cells that can be extracted from anindividual for therapeutic purposes, it is also necessary to have astimulation of cell proliferation. In a key publication, Waldman et al.reported, that in high-density 3D cultures low doses of CNP (10 to 100pM) elicited chondrocyte proliferation of up to 43% increase incellularity at the highest dose. Higher doses of CNP (10 nM)predominantly stimulated matrix deposition without affecting tissuecellularity (Waldman et al. 2008, Tissue Eng Part A. 14:441-448). CNP isthus suitable as a modulator of both chondrocyte proliferation and ECMdeposition during in vitro cartilage growth.

19. Tissue Engineering and Bone Regeneration, Especially for theAcceleration of Bone Healing or for the Improvement of Regenerating BoneTissue

The role of the NPR-B/CNP system as an important regulator of bonegrowth has been established by several publications: NPR-B knock-outmice displayed reduced bone growth (Tamura et al. 2004, Proc Natl AcadSci USA. 101:17300-17305, Pfeifer et al. 1996, Science. 274:2082-2086);mice with a deletion of the CNP gene also showed reduced bone growth,and this phenotype could be rescued by the overexpression of CNP inchondrocytes (Chusho et al. 2001, Proc Natl Acad Sci USA. 98:4016-4021);overexpression of BNP in mice resulted in skeletal overgrowth (Suda etal. 1998, Proc Natl Acad Sci USA. 95:2337-2342). More specifically, CNPwas able to promote chondrocyte proliferation and matrix formation(Krejci et al. 2005, J Cell Sci. 118:5089-5100, Ozasa et al. 2005, Bone.36:1056-1064). Using an organ culture of fetal mouse tibias, an in vitromodel of endochondral ossification, longitudinal bone growth wasstimulated by CNP (Yasoda et al. 1998, J Biol Chem. 273:11695-11700).

In summary, the experimental evidence strongly supports the use of CNPin bone regenerating applications.

20. Modulation of Neuronal Activity, Especially for Replacement of CNPin its “Central Nervous Function”

The extensive distribution of the NPR-C receptor in the brainstemsuggests an involvement of NPR-C in the neuromodulatory effect ofnatriuretic peptides (Abdelalim et al. 2008, Neuroscience. 155:192-202),which were shown to evoke a variety of peripheral effects when appliedto the brain (Puurunen and Ruskoaho 1987, Eur J Pharmacol. 141:493-495,Bianciotti et al. 2001, Regul Pept. 102:127-133).Intra-cerebroventricular administration of atrial natriuretic peptide inanaesthetized rats, for example, resulted in the stimulation of gastricacid secretion, that was totally abolished by vagotomy, suggesting vagusnerve involvement (Puurunen and Ruskoaho 1987, Eur J Pharmacol.141:493-495). In two studies by Sabbatini et al., the cerebroventricularadministration of CNP in rats dose-dependently enhanced the exocrinepancreatic fluid output through the activation of the NPR-C receptor andthe vago-vagal reflex (Sabbatini et al. 2005, Eur J Pharmacol.524:67-74, Sabbatini et al. 2007, Eur J Pharmacol. 577:192-202), thusmimicking the effect of endogenous CNP.

21. Cancer, Through Inhibition of Proliferation of Tumor Cells,Especially Glioma Cells, Neuroblastoma Cells, Adenocarcinoma Cells,Adenocarcinoma Cells in Breast Pancreas and Prostate, Melanoma Cells andRenal Carcinoma Cells

Several publications have shown the presence of natriuretic peptidereceptors on tumor cells, suggesting a potential to affect theproliferation of those cells via application of CNP, as has been shownin a range of other cell types.

Early in vitro data from cultured rat glioma cells demonstrated thepresence of receptors on those cells, that showed strongest activationby CNP, i.e. cGMP production (Eguchi et al. 1992, Eur J Pharmacol.225:79-82). In another cell line, a AtT-20 pituitary tumor cell line,the only natriuretic receptor present on the cell surface was the NPR-Breceptor. cGMP production in these AtT-20 cells was stimulated up to200-fold by CNP (Gilkes et al. 1994, Biochem J. 299 (Pt 2):481-487).

Western immunoblotting identified NPR-A and NPR-C receptors in humancolon adenocarcinoma cells. Application of 1 mM ANP to these cellsresulted in a decrease of up to 97% in cell number within 24 h,suggesting an anti-proliferative activity (Gower et al. 2005, Int JGastrointest Cancer. 36:77-87).

CNP caused a 39% decrease in the number of small-cell lung cancer cellsat 100 μM. The mechanism of growth inhibition supposedly is based on theinhibition of DNA synthesis, mediated in part by cGMP (Vesely et al.2005, Eur J Clin Invest. 35:388-398).

In yet another cell type, in human renal carcinoma cells, CNP alsodecreased the cell number, at a concentration of 100 μM by 10%. Thiseffect was sustained without any proliferation of the cells occurringfor three days after treatment with CNP. All three types of natriureticpeptide receptors, NPR-A, NPR-B, and NPR-C, were identified on renalcancer cells (Vesely et al. 2006, Eur J Clin Invest. 36:810-819).

22. Fibrosis, Especially Pulmonary Fibrosis, Renal Fibrosis, CardiacFibrosis, Hepatic Fibrosis or Systemic Fibrosis/Sclerosis

Several studies, investigating fibrotic events in different organsystems, have shown that the application of natriuretic peptides, inparticular of CNP, has a beneficial effect on disease progression. Amore general effect of CNP-mediated cGMP generation in fibroblasts isthe block of the activation of the mitogen-activated protein kinasecascade (Chrisman and Garbers 1999, J Biol Chem. 274:4293-4299), whichcould be exploited to treat any kind of fibrosis, in particular themultiorgan systemic fibrosis/sclerosis; treatment of single organfibrosis with CNP is supported by the following data:

In a model of bleomycin-induced pulmonary fibrosis in mice, infusion ofCNP markedly reduced bronchoalveolar lavage fluid levels of inflammatoryIL-1β, inhibited infiltration of macrophages into the alveolar andinterstitial regions, and markedly attenuated the fibrosis, as indicatedby significant decreases in Ashcroft score and lung hydroxyprolinecontent (Murakami et al. 2004, Am J Physiol Lung Cell Mol Physiol.287:L1172-1177).

With regard to kidney fibrosis, it was described that CNP had aninhibitory effect on the proliferation of glomerular mesangial cells(Suganami et al. 2001, J Am Soc Nephrol 12:2652-2663, Canaan-Kuhl et al.1998, Kidney Int 53:1143-1151, Osawa et al. 2000, Nephron. 86:467-472).In particular, CNP inhibited also MCP-1 secretion, and reduced collagenIV production from glomerular mesangial cells (Osawa et al. 2000,Nephron. 86:467-472).

Cardiac fibrosis, characterized by the proliferation of interstitialfibroblasts and the biosynthesis of extracellular matrix components inthe ventricles of the heart, is a consequence of remodeling processes.Soeki et al. showed that the application of CNP improved cardiacfunction and protected against cardiac remodeling after myocardialinfarct in rats (Soeki et al. 2005, J Am Coll Cardiol 45:608-616). Invitro, in cardiac fibroblasts, CNP had a suppressive effect onfibroblast proliferation and extracellular matrix production, the effectbeing stronger than by ANP or BNP (Horio et al. 2003, Endocrinology.144:2279-2284).

During chronic liver diseases, hepatic stellate cells, believed to playa role in the pathogenesis of liver fibrosis and portal hypertension(Friedman 1993, N Engl J Med. 328:1828-1835), acquired a myofibroblasticphenotype, proliferated, and synthesized components associated withfibrosis. The activation of NPR-B by CNP in myofibroblastic hepaticstellate cells was shown to inhibit both growth and contraction (Tao etal. 1999, J Biol Chem. 274:23761-23769), suggesting that during chronicliver diseases, CNP may counteract fibrogenesis.

C. Pharmaceutical Preparations

Other embodiments of the present invention are directed topharmaceutical compositions, comprising at least one novel NPR-B agonistdescribed herein, directed to the treatment or prevention of a diseasein a subject that is associated with elevated IOP, glaucoma, ocularhypertension, and/or retinal ganglion cell loss.

1. Effective Amount

As used herein, the term “effective amount,” or “therapeuticallyeffective amount,” refers to an amount of the agent that will activatethe function and/or activity of a type B natriuretic peptide receptor.The novel NPR-B agonists described herein lower intraocular pressure ortreat ocular hypertension in a patient having elevated IOP or ocularhypertension. Thus, an effective amount is an amount sufficient todetectably and repeatedly ameliorate, reduce, minimize or limit theextent of any disease associated with elevated intraocular pressure orocular hypertension, such as any of those diseases discussed above.

Treatment and/or prevention methods will involve treating an individualwith an effective amount of a composition containing a therapeuticallyeffective amount of at least one NPR-B agonist of the invention. Atherapeutically effective amount is described, generally, as that amountthat is known to be or suspected to be of benefit in the reduction ofthe signs or symptoms of a disease. In some embodiments of the presentinvention, an effective amount is generally an amount that is known orsuspected to be of benefit in reducing the signs or symptoms of glaucomaand associated optic nerve or retinal damage in a subject. It isenvisioned that the treatment with the NPR-B agonists hereof willstabilize or improve visual function (as measured by visual acuity,visual field, or other method known to those of ordinary skill in theart).

In some embodiments, an effective amount of a NPR-B agonist that may beadministered to a subject includes a dose from about 1 microgram/kg/bodyweight to about 500 microgram/kg/body weight or more per administration,and any range derivable therein.

2. Formulations

Regarding the methods set forth herein, a NPR-B agonist can beformulated in any manner known to those of ordinary skill in the art. Inthe compositions set forth herein, the concentration of a NPR-B agonistcan be any concentration known or suspected by those of ordinary skillin the art to be of benefit in the treatment and/or prevention ofophthalmic disease associated with elevated intraocular pressure orocular hypertension.

The actual dosage amount of a composition of the present inventionadministered to a subject can be determined by physical andphysiological factors such as body weight, severity of condition, thetype of disease being treated, previous or concurrent therapeuticinterventions, idiopathy of the patient and on the route ofadministration. The practitioner responsible for administration will, inany event, determine the concentration of active ingredient(s) in acomposition and appropriate dose(s) for the individual subject.

In certain non-limiting embodiments, the ophthalmic pharmaceuticalcompositions may comprise, for example, at least about 0.03%, by weightor volume, of an active ingredient. In other embodiments, the activeingredient may comprise between about 0.001% to about 75% of the weightor volume of the unit, or between about 0.01% to about 60%, and anyrange derivable therein. In more particular embodiments, thepharmaceutical composition may comprise between about 0.03% to about2.0% by weight or volume, of an active ingredient. In more particularembodiments, the composition comprises between about 0.05% to about 1.5%by weight or volume of an active ingredient. In further embodiments, thecomposition comprises between about 0.05% to about 1.2% by weight orvolume of an active ingredient.

A dose may be any amount of pharmaceutical composition that is known orsuspected to be of therapeutic benefit. For example, a dose may be about1 microgram/kg/body weight to about 500 microgram/kg/body weight or moreper administration, and any range derivable therein. A dose may berepeated as necessary as determined by one of ordinary skill in the artto achieve a desired therapeutic effect. For example, a dose may berepeated once, twice, three times, and so forth. In some embodiments, adose is administered twice a day, three times a day, four times a day,or more often. In further embodiments, a dose is administered everyother day, twice a week, once a month, or at a longer interval.

In certain embodiments of the present invention, the compositions setforth herein can include more than one NPR-B agonist. One of ordinaryskill in the art would be familiar with preparing and administeringpharmaceutical compositions that include more than one therapeuticagent. In some embodiments, the composition includes one or moreadditional therapeutic agents that are not NPR-B agonists.

In addition to the NPR-B agonists, the compositions of the presentinvention optionally comprise one or more excipients. Excipientscommonly used in pharmaceutical compositions include, but are notlimited to, carriers, tonicity agents, preservatives, chelating agents,buffering agents, surfactants and antioxidants.

A person of ordinary skill will recognize that the compositions of thepresent invention can include any number of combinations of ingredients(e.g., active agent, polymers, excipients, etc.). It is alsocontemplated that that the concentrations of these ingredients can vary.In non-limiting aspects, the percentage of each ingredient in thecomposition can be calculated by weight or volume of the totalcomposition. A person of ordinary skill in the art would understand thatthe concentrations can vary depending on the addition, substitution,and/or subtraction of ingredients in a given composition.

In some embodiments of the invention, a specific amount of a NPR-Bagonist is administered via the compositions described herein.

The phrase “pharmaceutically acceptable carrier” is art-recognized, andrefers to, for example, pharmaceutically acceptable materials,compositions or vehicles, such as a liquid or solid filler, diluent,excipient, solvent or encapsulating material, involved in carrying ortransporting any supplement or composition, or component thereof, fromone organ, or portion of the body, to another organ, or portion of thebody. Each carrier must be “acceptable” in the sense of being compatiblewith the other ingredients of the supplement and not injurious to thepatient.

Any of a variety of carriers may be used in the formulations of thepresent invention including water, mixtures of water and water-misciblesolvents, such as C1-7-alkanols, vegetable oils or mineral oilscomprising from 0.5 to 5% non-toxic water-soluble polymers, naturalproducts, such as gelatin, alginates, pectins, tragacanth, karaya gum,xanthan gum, carrageenin, agar and acacia, starch derivatives, such asstarch acetate and hydroxypropyl starch, and also other syntheticproducts, such as polyvinyl alcohol, polyvinylpyrrolidone, polyvinylmethyl ether, polyethylene oxide, preferably cross-linked polyacrylicacid, mixtures of those polymers. The concentration of the carrier is,typically, from 1 to 100000 times the concentration of the activeingredient.

Suitable tonicity-adjusting agents include mannitol, sodium chloride,glycerin, sorbitol and the like. Suitable preservatives includep-hydroxybenzoic acid ester, benzalkonium chloride, benzododeciniumbromide, polyquaternium-1 and the like. Suitable chelating agentsinclude sodium edetate and the like. Suitable buffering agents includephosphates, borates, citrates, acetates and the like. Suitablesurfactants include ionic and nonionic surfactants, though nonionicsurfactants are preferred, such as polysorbates, polyethoxylated castoroil derivatives and oxyethylated tertiary octylphenol formaldehydepolymer (tyloxapol). Suitable antioxidants include sulfites, ascorbates,BHA and BHT. The compositions of the present invention optionallycomprise an additional active agent.

In particular embodiments, the compositions are suitable for applicationto mammalian eyes. For example, for ophthalmic administration, theformulation may be a solution, a suspension, a gel, or an ointment.

In preferred aspects, the compositions that include NPR-B agonists willbe formulated for topical application to the eye in aqueous solution inthe form of drops. The term “aqueous” typically denotes an aqueouscomposition wherein the carrier is to an extent of >50%, morepreferably >75% and in particular >90% by weight water. These drops maybe delivered from a single dose ampoule which may preferably be sterileand thus rendering bacteriostatic or bacteriocidal components of theformulation unnecessary. Alternatively, the drops may be delivered froma multi-dose bottle which may preferably comprise a device whichextracts preservative from the formulation as it is delivered, suchdevices being known in the art.

In other aspects, components of the invention may be delivered to theeye as a concentrated gel or similar vehicle which forms dissolvableinserts that arc placed beneath the eyelids.

The compositions of the present invention may also be formulated assolutions that undergo a phase transition to a gel upon administrationto the eye.

In addition to the one or more NPR-B agonists, the compositions of thepresent invention may contain other ingredients as excipients. Forexample, the compositions may include one or more pharmaceuticallyacceptable buffering agents, preservatives (including preservativeadjuncts), non-ionic tonicity-adjusting agents, surfactants,solubilizing agents, stabilizing agents, comfort-enhancing agents,polymers, emollients, pH-adjusting agents and/or lubricants.

For topical formulations to the eye, the formulations are preferablyisotonic, or slightly hypotonic in order to combat any hypertonicity oftears caused by evaporation and/or disease. The compositions of thepresent invention generally have an osmolality in the range of 220-320mOsm/kg, and preferably have an osmolality in the range of 235-260mOsm/kg. The compositions of the invention have a pH in the range of5-9, preferably 6.5-7.5, and most preferably 6.9-7.4.

The formulations set forth herein may comprise one or morepreservatives. Examples of preservatives include quaternary ammoniumcompounds, such as benzalkonium chloride or benzoxonium chloride. Otherexamples of preservatives include alkyl-mercury salts of thiosalicylicacid, such as, for example, thiomersal, phenylmercuric nitrate,phenylmercuric acetate or phenylmercuric borate, sodium perborate,sodium chlorite, parabens, such as, for example, methylparaben orpropylparaben, alcohols, such as, for example, chlorobutanol, benzylalcohol or phenyl ethanol, guanidine derivatives, such as, for example,chlorohexidine or polyhexamethylene biguanide, sodium perborate, orsorbic acid.

In certain embodiments, the NPR-B agonists are formulated in acomposition that comprises one or more tear substitutes. A variety oftear substitutes are known in the art and include, but are not limitedto: monomeric polyols, such as, glycerol, propylene glycol, and ethyleneglycol; polymeric polyols such as polyethylene glycol; cellulose esterssuch hydroxypropylmethyl cellulose, carboxy methylcellulose sodium andhydroxy propylcellulose; dextrans such as dextran 70; water solubleproteins such as gelatin; vinyl polymers, such as polyvinyl alcohol,polyvinylpyrrolidone, and povidone; and carbomers, such as carbomer934P, carbomer 941, carbomer 940 and carbomer 974P. The formulation ofthe present invention may be used with contact lenses or otherophthalmic products.

In some embodiments, the compositions set forth herein have a viscosityof 0.5-10 cps, preferably 0.5-5 cps, and most preferably 1-2 cps. Thisrelatively low viscosity insures that the product is comfortable, doesnot cause blurring, and is easily processed during manufacturing,transfer and filling operations.

3. Route of Administration

Administration of the compositions of the invention can be by any methodknown to those of ordinary skill in the art, however, localadministration is preferred. It is contemplated that all local routes tothe eye may be used including topical, subconjunctival, periocular,retrobulbar, subtenon, intracameral, intravitreal, intraocular,subretinal, juxtascleral and suprachoroidal administration. Systemic orparenteral administration may be feasible including but not limited tointravenous, subcutaneous, intramuscular and oral delivery. The mostpreferred method of administration will be intravitreal or subtenoninjection of solutions or suspensions, or intravitreal or subtenonplacement of bioerodible or non-bioerodible devices, or by topicalocular administration of solutions or suspensions, or posteriorjuxtascleral administration of a gel formulation.

Those of skill in the art, in light of the present disclosure, willappreciate that obvious modifications of the embodiments disclosedherein can be made without departing from the spirit and scope of theinvention. All of the embodiments disclosed herein can be made andexecuted without undue experimentation in light of the presentdisclosure. The full scope of the invention is set out in the disclosureand equivalent embodiments thereof. The specification should not beconstrued to unduly narrow the full scope of protection to which thepresent invention is entitled.

While a particular embodiment of the invention has been shown anddescribed, numerous variations and alternate embodiments will occur tothose skilled in the art. Accordingly, the invention may be embodied inother specific forms without departing from its spirit or essentialcharacteristics. The described embodiments are to be considered in allrespects only as illustrative and not restrictive. The scope of theinvention is, therefore, indicated by the appended claims rather than bythe foregoing description. All changes to the claims that come withinthe meaning and range of equivalency of the claims are to be embracedwithin their scope. Further, all published documents, patents, andapplications mentioned herein are hereby incorporated by reference, asif presented in their entirety.

D. Secondary Forms of Therapy

In certain embodiments of the present invention, the subject isreceiving one or more secondary forms of therapy directed to treatmentor prevention of a particular eye disease.

A NPR-B agonist-containing ophthalmic composition of the presentinvention may be administered along with another agent or therapeuticmethod. For example, administration of the NPR-B agonist-containingcomposition of the present invention to a human subject may precede,follow, or be concurrent with other therapies for glaucoma, elevatedintraocular pressure or ocular hypertension. In some embodiments, theNPR-B agonist is formulated in the same composition as the secondaryform of therapy. In other embodiments, the NPR-B agonist is formulatedseparately from the secondary form of therapy. One of ordinary skill inthe art would be familiar with protocols for administering more than oneform of pharmacological therapy to a subject with a disease, and wouldbe familiar with methods of formulating more than one pharmacologicalagent in the same composition.

Examples of secondary therapeutic agents include, but are not limitedto: anti-glaucoma agents, such as beta-blockers including timolol,betaxolol, levobetaxolol, cartcolol, miotics including pilocarpine,carbonic anhydrase inhibitors, prostaglandins, seretonergics,muscarinics, dopaminergic agonists, adrenergic agonists includingapraclonidine and brimonidine; anti-angiogenesis agents; anti-infectiveagents including quinolones such as ciprofloxacin, and aminoglycosidessuch as tobramycin and gentamicin; non-steroidal and steroidalanti-inflammatory agents, such as suprofen, diclofenac, ketorolac,rimexolone and tetrahydrocortisol; growth factors, such as nerve growthfactor (NGF), basic fibroblast growth factor (bFGF), brain-derivedneurotrophic factor (BDNF), ciliary neutrophic factor (CNTF);immunosuppressant agents; and anti-allergic agents includingolopatadine. Information pertaining to olopatadine formulations can befound in U.S. Pat. No. 6,995,186, U.S. Patent App. Pub. No.2005/0158387, and U.S. Patent App. Pub. No. 2003/0055102, each of whichis hereby specifically incorporated by reference. The ophthalmic drugmay be present in the form of a pharmaceutically acceptable salt, suchas timolol maleate, brimonidine tartrate or sodium diclofenac.

Other examples of a secondary therapeutic agent include a receptortyrosine kinase (RTK) inhibitor. Exemplary RTK inhibitors are describedin U.S. Patent App. Pub. No. 2006/0189608, and U.S. Pat. No. 7,297,709,both of which are hereby specifically incorporated by reference. Inpreferred embodiments, the receptor tyrosine kinase inhibitor isN-[4-[3-amino-1H-indazol-4-yl]phenyl]-N′-(2-fluoro-5-methylphenyl)urea.

In other particular embodiments, the secondary therapeutic agent is aprostaglandin or a prostaglandin analog. For example, the prostaglandinanalog may be latanoprost, bimatoprost, unoprostone or travoprost.

In particular embodiments, the secondary therapeutic agent is a steroid.For example, the steroid may be a glucocorticoid, a progestin, amineralocorticoid, or a corticosteroid. Exemplary corticosteroidsinclude cortisone, hydrocortisone, prednisone, prednisolone,methylprednisone, triamcinolone, fluoromethalone, dexamethasone,medrysone, betamethasone, loteprednol, fluocinolone, flumethasone, ormometasone. Other examples of steroids include androgens, such astestosterone, methyltestosterone, or danazol. The secondary therapeuticagent may also be a glucocorticoid that is devoid of typicalglucocorticoid side-effects, such as a cortisene. Preferred cortisenesfor use in the methods of the invention include anecortave acetate andanecortave desacetate. Often steroids are administered as ester, acetal,or ketal prodrugs, many of which are water-insoluble. The secondarytherapeutic agents may be directed to treatment or prevention of asingle disease, or can be directed to treatment or prevention of two ormore diseases.

In addition to pharmacological agents, surgical procedures can beperformed in combination with the administration of the NPR-B agonists.One such surgical procedure can include laser trabeculoplasty ortrabeculectomy. In laser trabeculoplasty, energy from a laser is appliedto a number of noncontiguous spots in the trabecular meshwork. It isbelieved that the laser energy stimulates the metabolism of thetrabecular cells, and changes the extracellular material in thetrabecular meshwork.

Another surgical procedure may include filtering surgery. With filteringsurgery, a hole is made in the sclera near the angle. This hole allowsthe aqueous fluid to leave the eye through an alternate route. The mostcommonly performed filtering procedure is a trabeculectomy. In atrabeculectomy, a conjunctiva incision is made, the conjunctiva beingthe transparent tissue that covers the sclera. The conjunctiva is movedaside, exposing the sclera at the limbus. A partial thickness scleralflap is made and dissected half-thickness into the cornea. The anteriorchamber is entered beneath the scleral flap and a section of deep scleraand/or trabecular meshwork is excised. The scleral flap is loosely sewnback into place. The conjunctival incision is tightly closed.Post-operatively, the aqueous fluid passes through the hole, beneath thescleral flap which offers some resistance and collects in an elevatedspace beneath the conjunctiva called a bleb. The fluid then is eitherabsorbed through blood vessels in the conjunctiva or traverses acrossthe conjunctiva into the tear film.

E. Examples

The following examples are included to demonstrate preferred embodimentsof the invention. It should be appreciated by those of skill in the artthat the techniques disclosed in the examples which follow representtechniques discovered by the inventor to function well in the practiceof the invention, and thus can be considered to constitute preferredmodes for its practice. However, those of skill in the art should, inlight of the present disclosure, appreciate that many changes can bemade in the specific embodiments which are disclosed and still obtain alike or similar result without departing from the spirit and scope ofthe invention.

EXAMPLE 1

Material and Methods

The materials and methods as well as general methods are furtherillustrated by the following examples:

Solvents:

Solvents were used in the specified quality without furtherpurification.

Acetonitrile (Gradient grade, J.T. Baker); dichloromethane (forsynthesis, VWR); diethylether (for synthesis, VWR);N,N-dimethylformamide (LAB, VWR); dioxane (for synthesis, Aldrich);methanol (for synthesis, VWR).

Water: Milli-Q Plus, Millipore, demineralized.

Reagents:

The used reagents were purchased from Advanced ChemTech (Bamberg,Germany), Sigma-Aldrich-Fluka (Deisenhofen, Germany), Sachem(Heidelberg, Germany), J.T. Baker (Phillipsburg, USA), Iris Biotech(Marktredwitz, Germany), Lancaster (Griesheim, Germany), VWR (Darmstadt,Germany), NeoMPS (Strasbourg, France), Novabiochem (Bad Soden, Germany,from 2003 on Merck Biosciences, Darmstadt, Germany) and Acros (Geel,Belgium, distributor Fisher Scientific GmbH, Schwerte, Germany), Peptech(Cambridge, Mass., USA), Synthetech (Albany, Oreg., USA), Pharmacore(High Point, N.C., USA), Anaspec (San Jose, Calif., USA) and used in thespecified quality without further purification.

Non-commercially available non-conventional amino acids were preparedaccording to standard protocols either as building blocks for solidphase synthesis or by derivatization of commercially available aminoacids during solid phase synthesis.

If not stated differently, concentrations are given as percent byvolume.

Analysis of Peptides According to the Present Invention:

The analyses of peptides were performed with analytical HPLC methodsfollowed by either ESI-MS or MALDI-MS detection. For analyticchromatography a Hewlett Packard 1100-system together with an ESI-MS(Finnigan LCQ ion trap mass spectrometer) was used. Helium was used asimpact gas in the ion trap. For chromatographic separation aRP-18-column (Vydac (Merck) at 30° C. was used. A binary gradient wasapplied for all chromatograms (5-95% B, linear, A: 0.1% TFA in water andB: 0.1% TFA in CH3CN). UV detection was at λ=220 nm.

Analyses by means of HPLC/MS was performed using a linear gradient from95:5 to 5:95 (A: 0.1% TFA in water and B: 0.1% TFA in acetonitrile), RPcolumns were from the companies Phenomenex or Waters (Typ Luna C-18, 3μm, 2.00×50 mm, Symmetry C18 Column MV Kit, 5 μm, 4.6×250 mm,respectively); For ESI-MS measurements a mass spectrometerThermoFinnigan Advantage and/or LCQ Classic (both iontrap) was used. ForESI ionization helium served as impact gas in the ion trap. In case ofMALDI-MS analyses an Applied Biosystems Voyager RP MALDI massspectrometer was used with α-Cyano-4-hydroxycinnamic acid as internalcalibration matrix.

Purification of Peptides with Preparative HPLC:

Preparative HPLC separations were performed using Varian PLRP-S (10 μm,100 Å) columns (150×25 mm or 150×50 mm) with the following gradientsolvents: A: 0.05% TFA in H₂O and B: 0.05% TFA in CH₃CN

TABLE 4 Abbreviations: AAV general procedure Ac Acetyl AcmAcetamidomethyl DCM Dichloromethane DIC Diisopropylcarbodiimide DIPEAN,N-diisopropylethylamine DMF N,N-dimethylformamide DMSODimethylsulfoxide eq. Equivalent(s) ESI Electrospray ionisation FIG.Figure Fmoc 9-fluorenylmethyloxycarbonyl H hour(s) HATUO-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium-hexafluorophosphate HBTUO-(benzotriazol-1-yl)-1,1,3,3-tetramethyluronium- hexafluorophosphateHOBt 1-hydroxybenzotriazole HPLC high-pressure liquid chromatographyMALDI Matrix Assisted Laser Desorption/Ionization Me Methyl minminute(s) ml Milliliter MS Mass spectrometry MW Molecular weight NMPN-methylpyrrolidone Ph Phenyl RP Reversed phase ^(t)Bu tert-butyl THFTetrahydrofuran TIPS Triisopropyl silane TFA trifluoroacetic acid UVUltraviolet

EXAMPLE 2

Synthesis of Peptides

Linear peptides were synthesized using the Fmoc-tBu-strategy. Thesynthesis was done either manually in polypropylene syringes or via anautomatic synthesizer (Syro from Multisyntech, Witten or Sophas fromZinsser-Analytic, Frankfurt).

For the preparation of peptides carrying a C-terminal carboxylic acid,the C-terminal amino acid was either attached to a tritylchloride resin(approx. 100 mg resin; loading of reactive groups approx. 1.5 mmol/g;coupling with 0.8 eq. Fmoc-amino acid and 3.0 eq. DIPEA in DCM for 2 h;loading of the first amino acid approx. 0.2-0.4 mmol/g) or to Wang resin(100-200 mg resin; loading of reactive groups approx. 0.6 mmol/g;coupling with 4 eq. Fmoc-amino acid, 4 eq. DIC and 3 eq. NMI in DMF for3 h; loading of the first amino acid approx. 0.2-0.6 mmol/g).

For the preparation of peptides carrying a C-terminal carboxylic amide,the first amino acid was attached to the resin via Fmoc deprotection ofthe Fmoc-Rink amide resin (ca. 100 mg resin, ca. 0.5 mmol/g loading;Fmoc deprotection with 20% piperidine in DMF for 20 min) and subsequentcoupling of the Fmoc amino acid (reaction with 5 eq. Fmoc amino acid; 5eq. HBTU or 5 eq. HATU and 10 eq. DIPEA in NMP for 30-60 min and thisstep was optionally repeated).

After the coupling of the first amino acid, the synthesis of the peptidewas done via a repeated sequence of steps, as necessary, consisting ofFmoc deprotection and coupling of the corresponding Fmoc amino acid orcarboxylic acid. For the Fmoc deprotection the resin was treated with20% piperidine in DMF for 20 min. The coupling of the amino acids wascarried out via reaction with 5 eq. of the amino acid, 5 eq. HBTU or 5eq. HATU and 10 eq. DIPEA in DMF for 30-60 min. Each coupling step wasoptionally repeated.

For the introduction of the N-terminal acetyl group, the N-terminal freepeptide, bound to the resin, was incubated with a solution of 10% aceticacid anhydride and 20% DIPEA in DMF for 20 min. For the introduction ofthe N-terminal sulfonyl group, the N-terminal free peptide, bound to theresin, was incubated with a solution of 2 eq. of the correspondingsulfonyl chloride and 4 eq. DIPEA in DMF or DCM for 30 min and thistreatment was repeated once.

For the cleavage of the peptide from the resin and its side chainprotecting groups, a mixture of 95% TFA, 2.5% H2O, 2.5% TIPS or asimilar solution was added. Finally the crude peptide was isolatedeither by evaporation of TFA using a rotary evaporator or byprecipitation with the aid of methyl-^(t)butyl-ether at 0° C.

EXAMPLE 3

NPR-A Induced Production of Cyclic GMP in Stably Transfected Cell

To assess the specificity of compounds for NPR activation, human 293-Tcells transfected with NPR-A (Potter and Garbers 1992, J Biol Chem.267:14531-14534) are used in stimulation experiments.

In this homogenous assay, the cells are stimulated in suspension withthe test compound and the production of cyclic GMP (cGMP) is determined,from which EC50 values arc calculated. ANP, the naturally occurringligand of NPR-A is used as an internal control and to determine themaximal cGMP production of the cells, which allows the calculation ofactivation values of the tested compounds relative to ANP.

Preparation of cells: NPR-A transfected 293-T cells are washed once withphosphate buffered saline (PBS) and detached from a 75 cm² tissueculture flask by addition of 3 ml of non enzymatic cell dissociationsolution (Sigma-Aldrich) and incubation for 10 min. at room temperature.Detached cells are harvested in 20 ml PBS and centrifuged for 10 min at200×g at room temperature. The cells are resuspended in DMEM/Ham's F12mix supplemented with 1 mM IBMX (Medium) and adjusted to a density of1.25×10⁵ cells/ml and incubated for 15 min. at room temperature.

Stimulation of cells: 20 μl of cells (2.5×10³ cells) arc added to eachwell of a 96 well white optical bottom tissue culture plate (Nunc,Germany). 10 μl of compound dilution is added and the cells arestimulated for 25 min. at room temperature. The stimulation is stoppedby addition of 20 μl of Lysis buffer (reagent included in cGMP AssayKit).

Determination of cGMP: The amount of produced cGMP in the cells isdetermined using HitHunter™ cGMP Assay kit (DiscoveRX) according tomanufacturer's instructions.

Dilution of compounds: For EC50 determinations, duplicate wells arestimulated with a serial dilution of a 10 mM DMSO compound stocksolution. Dilutions are prepared in Medium supplemented with IBMX (1mM). The final compound concentration in the assay is in the range from45 μM to 20 nM. The internal standard compound ANP is used atconcentrations ranging from 5 μM to 310 pM.

EXAMPLE 4

NPR-B Induced Production of Cyclic GMP in Human Glaucoma TrabecularMeshwork Cells (GTM-3)

The potency of compounds to activate NPR-B was evaluated in a functionalassay using endogenously NPR-B expressing GTM-3 cells (Pang, Shade etal. 1994). In this assay the dose dependent production of cyclic GMP(cGMP) is determined and EC₅₀ values are calculated. The naturaloccurring ligand for NPR-B, i.e. CNP is used as an internal control andto determine the maximal cGMP production of the cells, which allows thecalculation of activation values of the tested compounds relative toCNP.

Preparation of cells: In a 96 well white optical bottom tissue cultureplate (Nunc, Germany) 1.5×10⁵ cells/well are seeded in Dulbecco's MEM(DMEM, Biochrom) supplemented with Gentamycin (0.056 mg/ml) andincubated for 18 h with 10% CO₂ in a humidified atmosphere.

Stimulation of cells: The cell culture medium is aspirated and each wellis washed with 200 μl DMEM/Ham's F12=Medium (Gibco). Then, 200 μl Mediumsupplemented with 1.5 mM IBMX (3-Isobutyl-1-methyl-Xanthin, Sigma) isadded to each well and incubated for 15 min. at room temperature. 25 μlof compound dilution is added and the cells are stimulated for 15 min.at room temperature. The stimulation is stopped by aspiration of themedium and addition of 20 μl of Lysis buffer (reagent included in cGMPAssay Kit).

Determination of cGMP: The amount of produced cGMP in the cells isdetermined using HitHunter™ cGMP Assay kit (DiscoveRX) according tomanufacturer's instructions.

Dilution of compounds: For EC₅₀ determinations, duplicate wells arestimulated with a serial dilution of a 10 mM DMSO compound stocksolution. Dilutions are prepared in Medium supplemented with IBMX (1.5mM). Final compound concentrations are in the range from 45 μM to 20 nM.Highly active compounds, e.g. CNP are used for stimulation atconcentrations ranging from 5μM to 6 nM.

EXAMPLE 5

Efficacy in the Rabbit

A single 30 μL drop of a test item formulation was administered torabbit eyes (n=8 to 10).

Intraocular pressure (IOP) was assessed in each eye at 0 hr, just priorto dosing, and again hourly for up to 4 hr post dose. The efficacy of agiven formulation was determined based on the difference between thepretreatment TOP readings at 0 hr and the post treatment readings. Amaximum percent reduction in IOP greater than 15% was noted by the “+”symbol. A maximum IOP reduction of less than 15% was assigned the “−”symbol.

Results obtained with novel compounds of the invention in theabove-described assays are provided in Table 5, below:

TABLE 5 In vivo results with novel compounds of the invention accordingto the methods described in Example 5. RIOP SEQ dose 300 ug ID − IOPreduction <15% NO: JAL STRUCTURE + IOP reduction >15% 3 CNP CNP − 81 781⁺ Occ-ala-ala-Phe-Gly-Leu-Pro-Leu-Asp-Arg- − Lle-NH₂; 127  955⁺⁺Occ-pro-Phe-Gly-Leu-Pro-Nml-Asp-Arg-Ile- − NH₂; 130  958⁺⁺Occ-Sni-Nmf-Gly-Leu-Pro-Nml-Asp-Arg-Ile- − NH₂; 135  967⁺Occ-Sni-Nmf-Gly-Leu-Pro-Leu-Asp-Arg-Ile- − NH₂; 182 1041⁺Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- + NH₂; 203 1085⁺Occ-ala-Nmf-arg-Leu-Hyp-Nml-Asp-Arg-Ile- + NH₂; 187 1047⁺⁺Occ-ala-Phe-arg-Leu-Hyp-Leu-Asp-Arg-Ile- + NH₂; 204 1086⁺⁺Occ-ala-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile- + NH₂; 183 1042⁺Occ-ala-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- − NH₂; 195 1060⁺⁺Occ-ala-Phe-lys-Leu-Hyp-Nml-Asp-Arg-Ile- + NH₂; 267 1287Occ-Sni-Phe-dap(Me2)-Leu-Hyp-Nml-Asp- + Arg-Ile-NH₂; 274 1295⁺Occ-Sni-Phe-leu-Leu-Tap-Nml-Asp-Arg-Ile- + NH₂; 355 1400⁺Occ-Sni-Phe-dap(Me2)-Leu-Hyp-Nml-Val-Arg- + Ile-NH₂; 292 1325⁺Occ-Sni-Phe-dap(Me2)-Leu-Tap-Nml-Asp-Arg- + Ile-NH₂; 332 1369⁺Oct-Sni-Phe-dap(Me2)-Leu-Tap-Nml-Asp-Arg- + Ile-NH₂; 372 1429⁺⁺Oct-Sni-Phe-dap(Me2)-Leu-Hyp-Nml-Asp-Arg- + Ile-NH₂; 414 1496+Occ-Sni-Eaa-leu-Leu-Hyp-Nml-Asp-Arg-Ile- − NH₂; 421 1512++Occ-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Pro-Che; + 425 1555++Occ-Sni-Phe-Apc-Leu-Hyp-Nml-Asp-Arg-Ile- + NH₂; 481 1654+Occ-Sni-Phe-leu-Leu-Tap-Nml-Val-Pro-Che; − 506 1729+Occ-Sni-Phe-leu-Leu-Tap-Nml-Val-Arg-Che; + 507 1730+Occ-Sni-Phe-leu-Leu-Hyp-Nml-Val-Arg-Che; + 269 1289+Occ-Sni-Phe-orn(Me2)-Leu-Hyp-Nml-Asp- + Arg-Ile-NH₂ HCl salt except*TFA; Dose is 300 μg topical ocular unless (##); DB rabbits unless NZA,scores 1-4 (4 = IOP could not be taken); “” indicates hypertensivephase; (n = #R) means # of responders out of 10-12 animals tested; 1% is+ susp ++ sol

* * *

All of the methods disclosed and claimed herein can be made and executedwithout undue experimentation in light of the present disclosure. Whilethe methods of this invention have been described in terms of preferredembodiments, it will be apparent to those of skill in the art thatvariations may be applied to the methods described herein withoutdeparting from the concept, spirit, and scope of the invention. Morespecifically, it will be apparent that certain agents which are bothchemically and physiologically related may be substituted for the agentsdescribed herein while the same or similar results would be achieved.All such similar substitutes and modifications apparent to those skilledin the art are deemed to be within the spirit, scope, and concept of theinvention as defined by the appended claims.

All references cited herein, to the extent that they provide exemplaryprocedural or other details supplementary to those set forth herein, arespecifically incorporated herein by reference.

What is claimed is:
 1. A method for treating ophthalmic disease in apatient in need thereof, said method comprising administering to saidpatient a therapeutically effective amount of a compound having thefollowing formula:

wherein B is selected from the group consisting of R^(b1)— andR^(b2)—C(O)—, wherein: R^(b1) is selected from the group consisting ofC₆-C₁₀ alkyl and C₆-C₁₀ alkyl substituted by NR^(b4)R^(b5); R^(b2) isC₆-C₁₀ alkyl substituted by NR^(b4)R^(b5); and R^(b4) and R^(b5) are,independently, selected from the group consisting of H and C₁-C₄ alkyl;and R^(11b) is selected from the group consisting of H, C₁-C₈ alkyl,C₄-C₈ cycloalkyl, C₇-C₁₂ bicycloalkyl, and C₁-C₄ alkyl-C₄-C₈ cycloalkyl.2. The method of claim 1, wherein: R^(11b) is selected from the groupconsisting of H and C₁-C₈ alkyl.
 3. The method of claim 1, wherein saidophthalmic disease is glaucoma, elevated intraocular pressure, or ocularhypertension.
 4. A method for treating ophthalmic disease in a patientin need thereof, said method comprising administering to said patient atherapeutically effective amount of a compound having the followingformula:

wherein B is selected from the group consisting of R^(b1)— andR^(b2)—C(O)—, wherein: R^(b1) is selected from the group consisting ofC₆-C₁₀ alkyl and C₆-C₁₀ alkyl substituted by NR^(b4)R^(b5); R^(b2) is C₆-C₁₀ alkyl substituted by NR^(b4)R^(b5); and R^(b4) and R^(b5) are,independently, selected from the group consisting of H and C₁-C₄ alkyl;and R^(11b) is selected from the group consisting of H, C₁-C₈ alkyl,C₄-C₈cycloalkyl, C₇-C₁₂ bicycloalkyl, and C₁-C₄alkyl-C₄-C₈cycloalkyl. 5.The method of claim 4, wherein: R^(11b) is selected from the groupconsisting of H and C₁-C₈ alkyl.
 6. The method of claim 4, wherein saidophthalmic disease is glaucoma, elevated intraocular pressure, or ocularhypertension.
 7. A method for treating ophthalmic disease in a patientin need thereof, said method comprising administering to said patient atherapeutically effective amount of a compound selected from:(SEQ ID NO: 228) 6054-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 244) Occ-Sni-Mpa-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 245) Occ-Sni-Ppa-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 246) (6071-OH)-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 248) 5587-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 270) (AR-201-48)-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 272) (AR-201-48)-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 307) (AR-201-58)-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 308) (AR-201-59)-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 311) Sbt-Sni-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 313) Sbt-ala-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 314) Nbt-ala-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 339) (AR-201-72)-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 340) (AR-201-72)-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 374) Miy-Gab-Hgl-ala-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile- NH2;(SEQ ID NO: 375) Ac-Miy-Gab-Hgl-ala-Phe-arg-Leu-Hyp-Nml-Asp-Arg-Ile-NH2; (SEQ ID NO: 408) Occ-Sni-Tyr-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 409) Occ-Sni-Bmf-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 423) Occ-Sni-Phe-Ala-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 437) Occ-Sni-Phe-nml-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 457) 2553-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 460) 6988-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 462) 1695-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 467) Occ-Sni-Otf-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 474) Occ-Sni-NHfe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 484) 785-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 486) 3218-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 489) 1281-G-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 490) 1281-Bal-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 504) Occ-Sni-Hfe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 505) Occ-ala-Nmf-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 533) Gluc-Aoa-hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- NH2;(SEQ ID NO: 535) (1270)-Hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 537) Occ-Hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 555) (AR-314-170)-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 556) (AR-314-171)-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2;(SEQ ID NO: 557) (AR-385-008)-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH2; and(SEQ ID NO: 565) Occ-Sni-Phe-thr-Leu-Hyp-Nml-Asp-Arg-Ile-NH2.


8. The method of claim 7, wherein said ophthalmic disease is glaucoma,elevated intraocular pressure, or ocular hypertension.
 9. The method ofclaim 7, wherein the compound is selected from: (SEQ ID NO: 457)2553-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; (SEQ ID NO: 460)6988-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; (SEQ ID NO: 462)1695-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; (SEQ ID NO: 484)785-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; (SEQ ID NO: 486)3218-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; (SEQ ID NO: 489)1281-G-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; (SEQ ID NO: 490)1281-Bal-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂; (SEQ ID NO: 533)Gluc-Aoa-hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- NH₂; (SEQ ID NO: 535)(1270)-Hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile- NH₂; and(SEQ ID NO: 537) Occ-Hgl-Sni-Phe-leu-Leu-Hyp-Nml-Asp-Arg-Ile-NH₂.


10. The method of claim 9, wherein said ophthalmic disease is glaucoma,elevated intraocular pressure, or ocular hypertension.